Bicyclic modulators of androgen receptor function

ABSTRACT

The invention provides compounds according to formula I 
     
       
         
         
             
             
         
       
         
         
           
             wherein the substitutents are as described herein. 
           
         
       
    
     Further provided are methods of using such compounds for the treatment of nuclear hormone receptor-associated conditions, such as age related diseases, for example sarcopenia. Also provided are pharmaceutical compositions containing such compounds and processes for preparing some of the compounds of the invention.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a Continuation-in-Part Application of U.S. patentapplication Ser. No. 10/438,722, filed May 15, 2003, now abandoned thatclaims the benefit of U.S. Provisional Application Nos. 60/381,616,filed May 17, 2002 and 60/406,711, filed Aug. 29, 2002, which areincorporated herein by reference in their entirety.

FIELD OF THE INVENTION

The present invention relates to bicyclic compounds, methods of usingsuch compounds in the treatment of androgen receptor-associatedconditions, such as age-related diseases, for example sarcopenia, and topharmaceutical compositions containing such compounds.

BACKGROUND OF THE INVENTION

Nuclear hormone receptors (NHR's) constitute a large super-family ofstructurally-related and sequence-specific gene regulators scientistshave named “ligand-dependent transcription factors.” R. M. Evans,Science, 240:889 (1988). The steroid binding NHR's (SB—NHR's) form arecognized subset of the NHR's, including the progesterone receptor(PR), androgen receptor (AR), estrogen receptor (ER), glucocorticoidreceptor (GR) and mineralocorticoid receptor (MR). The conventionalnuclear hormone receptors are generally transactivators in the presenceof ligand, which selectively bind to the NHR in a way that effects genetranscription. In the absence of a corresponding ligand, some of theorphan receptors behave as if they are transcriptionally inert. Others,however, behave as either constitutive activators or repressors. Theseorphan nuclear hormone receptors are either under the control ofubiquitous ligands that have not been identified, or do not need to bindligand to exert these activities.

The AR is a ligand-activated transcriptional regulatory protein thatmediates induction of male sexual development and function through itsactivity with endogenous androgens. In addition, androgens areassociated with male and female maintenance of muscle mass and strength,bone mass and erythropoiesis. Androgens, such as testosterone, also playan important role in many physiological processes, such asdifferentiation of male internal and external genitalia, development andmaintenance of male secondary sexual characteristics (e.g., thedevelopment of prostate, seminal vesicles, penis, scrotum, skeletalmuscle, redistribution of body fat, stimulation of long bone growth,closure of epiphyses, development of male hair growth pattern andenlargement of larynx), the maintenance of sexual behavior and function(e.g., libido and potency) and spermatogenesis (in man).

As one ages, the serum androgen concentration in the body declines. Theage dependent decline in androgens is associated with changes in bodycomposition for men and women, such as a lower percentage of muscle massand an increase in body fat, e.g., sarcopenia. In this regard,modulation of the AR gene can have an impact on the physiologicaleffects associated with androgen production. However, the effectivenessof known modulators of steroid receptors is often tempered by theirundesired side-effect profile, particularly during long-termadministration. For example, the administration of synthetic androgenshas been associated with liver damage, prostate cancer, adverse effectson male sexual function and adverse effects associated withcardiovascular and erythropoietic function.

Numerous synthetically-derived steroidal and non-steroidal agonists andantagonists have been described for the members of the SB-NHR family.Many of these agonist and antagonist ligands are used clinically in manto treat a variety of medical conditions. RU486 (mifepristone) is anexample of a synthetic antagonist of the PR, which is utilized as abirth control agent (Vegeto et al., Cell 69: 703-713 (1992)). Flutamideis an example of an antagonist of the AR, which is utilized for thetreatment of prostate cancer (Neri et al, Endo. 91, 427-437 (1972)).Tamoxifen is an example of a tissue-selective modulator of the ERfunction, that is used in the treatment of breast cancer (Smigel J.Natl. Cancer Inst. 90, 647-648 (1998)). Tamoxifen can function as anantagonist of the ER in breast tissue while acting as an agonist of theER in bone (Grese et al., Proc. Natl. Acad. Sci. USA 94, 14105-14110(1997)). Because of the tissue-selective effects seen for Tamoxifen,this agent, and agents like it, are referred to as tissue-selectiveestrogen receptor modulators. In addition to synthetically-derivednon-endogenous ligands, non-endogenous ligands for NHR's can be obtainedfrom food sources (Regal et al., Proc. Soc. Exp. Biol. Med. 223, 372-378(2000) and Hempstock et al., J. Med. Food 2, 267-269 (1999)). Theflavanoid phytoestrogens are an example of an unnatural ligand forSB-NHR's that are readily obtained from a food source such as soy(Quella et al., J. Clin. Oncol. 18, 1068-1074 (2000) and Banz et al., J.Med. Food 2, 271-273 (1999)). The ability to modulate thetranscriptional activity of an individual NHR by the addition of a smallmolecule ligand, makes these receptors ideal targets for the developmentof pharmaceutical agents for a variety of disease states.

As mentioned above, non-natural ligands can be synthetically engineeredto serve as modulators of the function of NHR's. In the case ofSB-NHR's, engineering of an unnatural ligand can include theidentification of a core structure which mimics the natural steroid coresystem. This can be achieved by random screening against severalSB-NHR's, or through directed approaches using the available crystalstructures of a variety of NHR ligand binding domains (Bourguet et al.,Nature 375, 377-382 (1995), Brzozowski, et al., Nature 389, 753-758(1997), Shiau et al., Cell 95, 927-937 (1998) and Tanenbaum et al.,Proc. Natl. Acad. Sci. USA 95, 5998-6003 (1998)). Differentialsubstitution about such a steroid mimic core can provide agents withselectivity for one receptor versus another. In addition, suchmodifications can be employed to obtain agents with agonist orantagonist activity for a particular SB-NHR. Differential substitutionabout the steroid mimic core can result in the formation of a series ofhigh affinity agonists and antagonists with specificity for, forexample, ER versus PR versus AR versus GR versus MR. Such an approach ofdifferential substitution has been reported, for example, for quinolinebased modulators of steroid NHRs in Hamann et. al., J. Med. Chem., 41,623 (1998); Hamann et. al., J. Med. Chem. 42, 210 (1999); WO 9749709;U.S. Pat. No. 5,696,133; U.S. Pat. No. 5,696,130; U.S. Pat. No.5,696,127; U.S. Pat. No. 5,693,647; U.S. Pat. No. 5,693,646; U.S. Pat.No. 5,688,810; U.S. Pat. No. 5,688,808 and WO 9619458, all incorporatedherein by reference.

Accordingly, identification of compounds which have good specificity forone or more steroid receptors, but which have reduced or nocross-reactivity for other steroid or intracellular receptors, would beof significant value in the treatment of male and femalehormone-responsive diseases. There is, therefore, a need in the art forthe identification of selective modulators of the steroid bindingnuclear hormone receptors, particularly non-steroidal, non-toxic tissueselective androgen receptor modulators, which activate the androgenreceptor in skeletal muscle while demonstrating limited or neutraleffect on other androgen responsive (e.g., prostate) tissues.

SUMMARY OF THE INVENTION

In accordance with illustrative embodiments and demonstrating featuresof the present invention, compounds are provided which are capable ofmodulating the function of a nuclear hormone receptor. Preferably thecompounds are selective androgen receptor modulators, and have thegeneral formula I

wherein

R₁ is selected from the group consisting of hydrogen (H), alkyl orsubstituted alkyl, alkenyl or substituted alkenyl, arylalkyl orsubstituted arylalkyl, CO₂R₄, CONR₄R₄′ and CH₂OR₄;

R₂ and R₂′ are independently selected from the group consisting ofhydrogen (H), OR₃, SR₃, halo, NHR₃, NHCOR₄, NHCO₂R₄, NHCONR₄R₄′ andNHSO₂R₄;

R₃ in each functional group is independently selected from the groupconsisting of hydrogen (H), alkyl or substituted alkyl, CHF₂, CF₃ andCOR₄;

R₄ and R₄′ in each functional group are each independently selected fromthe group consisting of hydrogen(H), alkyl or substituted alkyl, alkenylor substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl orsubstituted cycloalkyl, arylalkyl or substituted arylalkyl, aryl orsubstituted aryl, and heteroaryl or substituted heteroaryl;

R₅ and R₅′ are each independently selected from the group consisting ofhydrogen(H), alkyl or substituted alkyl, alkenyl or substituted alkenyl,alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl,arylalkyl or substituted arylalkyl, aryl or substituted aryl, andheteroaryl or substituted heteroaryl, wherein at least one of R₅ and R₅′is hydrogen, or R₅ and R₅′ taken together can form a double bond withoxygen (O), sulfur (S), NR₇ or CR₇R₇′;

R₆ and R₆′ are each independently selected from the group consisting ofhydrogen(H), alkyl or substituted alkyl, alkenyl or substituted alkenyl,alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl,arylalkyl or substituted arylalkyl, aryl or substituted aryl, andheteroaryl or substituted heteroaryl, wherein at least one of R₆ and R₆′is hydrogen, or R₆ and R₆′ taken together can form a double bond withoxygen (O), sulfur (S), NR₇ or CR₇R₇′;

R₇ and R₇′ in each functional group are each independently selected fromthe group consisting of hydrogen(H), OR₄, alkyl or substituted alkyl,alkenyl or substituted alkenyl, alkynyl or substituted alkynyl,cycloalkyl or substituted cycloalkyl, arylalkyl or substitutedarylalkyl, aryl or substituted aryl and heteroaryl or substitutedheteroaryl

G is an aryl, heterocyclo or heteroaryl group, wherein said group ismono- or polycyclic, and which is optionally substituted with one ormore substituents selected from the group consisting of hydrogen, halo,CN, CF₃, OR₄, CO₂R₄, NR₄R₄′, CONR₄R₄′, CH₂OR₄, alkyl or substitutedalkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl,cycloalkyl or substituted cycloalkyl, arylalkyl or substitutedarylalkyl, aryl or substituted aryl and heteroaryl or substitutedheteroaryl;

W is selected from the group consisting of (CR₆R₆′), C(R₆)OR₃,C(R₆)(NR₄R₄′), and

n is an integer of 1 or 2.

The definition of formula I above includes of all prodrug esters,stereoisomers and pharmaceutically acceptable salts of formula I.

Further embodiments of the present invention include compounds of theformula Ih

wherein

R₁ is selected from the group consisting of hydrogen (H), alkyl orsubstituted alkyl, alkenyl or substituted alkenyl, arylalkyl orsubstituted arylalkyl, CO₂R₄, CONR₄R₄′ and CH₂OR₄;

R₂ and R₂′ are each independently selected from the group consisting ofhydrogen (H), OR₃, SR₃, halo, NHR₃, NHCOR₄, NHCO₂R₄, NHCONRR′ andNHSO₂R₄;

R₃ in each functional group is independently selected from the groupconsisting of hydrogen (H), alkyl or substituted alkyl, CHF₂, CF₃ andCOR₄;

R₄ and R₄′ in each functional group are each independently selected fromthe group consisting of hydrogen(H), alkyl or substituted alkyl, alkenylor substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl orsubstituted cycloalkyl, arylalkyl or substituted arylalkyl, aryl orsubstituted aryl, and heteroaryl or substituted heteroaryl;

R₆ and R₆′ are each independently selected from the group consisting ofhydrogen(H), alkyl or substituted alkyl, alkenyl or substituted alkenyl,alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl,arylalkyl or substituted arylalkyl, aryl or substituted aryl, andheteroaryl or substituted heteroaryl, wherein at least one of R₆ and R₆′is hydrogen, or R₆ and R₆′ taken together can form a double bond withoxygen (O), sulfur (S), NR₇ or CR₇R₇′;

X and Y are each independently oxygen (O) or sulfur (S);

G is an aryl, heterocyclo or heteroaryl group, wherein said group ismono- or polycyclic, and which is optionally substituted with one ormore substitutents selected from the group consisting of hydrogen, halo,CN, CF₃, OR₄, CO₂R₄, NR₄R₄′, CONR₄R₄′, CH₂OR₄, alkyl or substitutedalkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl,cycloalkyl or substituted cycloalkyl, arylalkyl or substitutedarylalkyl, aryl or substituted aryl and heteroaryl or substitutedheteroaryl; and

W is selected from the group consisting of (CR₆R₆′), C(R₆)OR₃,C(R₆)(NR₄R₄′), and

n is an integer of 1 or 2.

The definition of formula Ih above is inclusive of all prodrug esters,stereoisomers and pharmaceutically acceptable salts of formula Ih.

The compounds of formula I and formula Ih modulate the function of thenuclear hormone receptors, particularly the androgen receptor, andinclude compounds which are, for example, selective agonists, partialagonists, antagonists or partial antagonists of the androgen receptor.Preferably the compounds of formula I possess activity as agonists ofthe androgen receptor and may be used in the treatment of diseases ordisorders associated with androgen receptor activity, such asmaintenance of muscle strength and function (e.g., in the elderly);reversal or prevention of frailty or age-related functional decline(“ARFD”) in the elderly (e.g., sarcopenia); prevention of catabolic sideeffects of glucocorticoids; prevention and treatment of reduced bonedensity or growth (e.g., osteoporosis and osteopenia); treatment ofchronic fatigue syndrome (CFS); chronic myalgia; treatment of acutefatigue syndrome and muscle loss following elective surgery (e.g.,post-surgical rehabilitation); acceleration of wound healing;accelerating bone fracture repair (such as accelerating the recovery ofhip fracture patients); treatment of wasting secondary to fractures andwasting in connection with chronic obstructive pulmonary disease (COPD),chronic liver disease, AIDS, weightlessness, cancer cachexia, burn andtrauma recovery, chronic catabolic state (e.g., coma), eating disorders(e.g., anorexia) and chemotherapy.

The present invention provides for compounds of formula I and Ih,pharmaceutical compositions employing such compounds and for methods ofusing such compounds. In particular, the present invention provides apharmaceutical composition comprising a therapeutically effective amountof a compound of formula I, Ih or both, alone or in combination with apharmaceutically acceptable carrier.

Further, in accordance with the present invention, a method is providedfor preventing, inhibiting or treating the progression or onset ofdiseases or disorders associated with nuclear hormone receptors,particularly, the androgen receptor, such as the diseases or disordersdefined above and hereinafter, wherein a therapeutically effectiveamount of a compound of formula I, Ih or both, is administered to amammalian, i.e., human, patient in need of treatment.

The compounds of the invention can be used alone, in combination withother compounds of the present invention, or in combination with one ormore other agent(s) active in the therapeutic areas described herein.

In addition, a method is provided for preventing, inhibiting or treatingthe diseases as defined above and hereinafter, wherein a therapeuticallyeffective amount of a combination of a compound of formula I, Ih orboth, and another type of therapeutic agent, is administered to a humanpatient in need of treatment.

Preferred are compounds of formula I where R₅ and R₅′ are hydrogen orare taken together form a double bond with oxygen (O) or sulfur (S); and

R₆ and R₆′ are taken together form a double bond with oxygen (O) orsulfur (S).

Additional preferred embodiments include are compounds of formula I andIh wherein

R₁ is hydrogen (H) or alkyl; and

R₂ is hydroxyl (OH).

Further preferred embodiments include compounds of formula I and Ihwhere G is selected from:

wherein

R₈, R₉, R₁₀ and R₁₁ in each functional group are each independentlyselected from the group consisting of hydrogen (H), NO₂, CN, CF₃, OR₄,CO₂R₄, NR₄R₄′, CONR₄R₄′, CH₂OR₄, alkyl or substituted alkyl, alkenyl orsubstituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl orsubstituted cycloalkyl, arylalkyl or substituted arylalkyl, aryl orsubstituted aryl and heteroaryl or substituted heteroaryl;

A to F is each independently selected from N or CR₁;

J, K, L, P and Q are each independently selected from NR₁₂, O, S, SO,SO₂ or CR₁₂R₁₂;

R₁₂ and R₁₂′ in each functional group are each independently selectedfrom a bond or R₁; and

m is an integer of 0 or 1.

Preferred are compounds of formula I and Ih where R₈ is CN.

The present invention also provides processes for preparing somecompounds of the invention.

DETAILED DESCRIPTION OF THE INVENTION

Thus, in a first embodiment, the present invention provides compoundsaccording to formula I

wherein

R₁ is selected from hydrogen (H), alkyl or substituted alkyl, alkenyl orsubstituted alkenyl, arylalkyl or substituted arylalkyl, CO₂R₄, CONR₄R₄′and CH₂OR₄;

R₂ and R₂′ are each independently selected from hydrogen (H), alkyl,substituted alkyl, OR₃, SR₃, halo, NHR₄, NHCOR₄, NHCO₂R₄, NHCONR₄R₄′ andNHSO₂R₄;

and at least one of R₂ and R₂′ is H or alkyl, with the exception that R₂and R₂′ can both be OR₃ when R₃ is not H;

R₃ in each functional group is independently selected from hydrogen (H),alkyl or substituted alkyl, CHF₂, CF₃ and COR₄;

R₄ and R₄′ in each functional group are each independently selected fromhydrogen(H), alkyl or substituted alkyl, alkenyl or substituted alkenyl,alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl,arylalkyl or substituted arylalkyl, aryl or substituted aryl, andheteroaryl or substituted heteroaryl;

R₅ and R₅′ are each independently selected from hydrogen(H), alkyl orsubstituted alkyl, alkenyl or substituted alkenyl, alkynyl orsubstituted alkynyl, cycloalkyl or substituted cycloalkyl, arylalkyl orsubstituted arylalkyl, aryl or substituted aryl, and heteroaryl orsubstituted heteroaryl, wherein at least one of R₅ and R₅′ is hydrogen,or R₅ and R₅′ taken together can form a double bond with oxygen (O),sulfur (S), NR₇ or CR₇R₇′;

R₆ and R₆′ are each independently selected from hydrogen(H), alkyl orsubstituted alkyl, alkenyl or substituted alkenyl, alkynyl orsubstituted alkynyl, cycloalkyl or substituted cycloalkyl, arylalkyl orsubstituted arylalkyl, aryl or substituted aryl, and heteroaryl orsubstituted heteroaryl, wherein at least one of R₆ and R₆′ is hydrogen,or R₆ and R₆′ taken together can form a double bond with oxygen (O),sulfur (S), NR₇ or CR₇R₇′;

R₇ and R₇′ in each functional group are each independently selected fromhydrogen(H), OR₄, alkyl or substituted alkyl, alkenyl or substitutedalkenyl, alkynyl or substituted alkynyl, cycloalkyl or substitutedcycloalkyl, arylalkyl or substituted arylalkyl, aryl or substituted aryland heteroaryl or substituted heteroaryl;

G is an aryl, heterocyclo or heteroaryl group, wherein said group ismono- or polycyclic, and which is optionally substituted with one ormore substitutents selected from hydrogen, halo, CN, CF₃, OR₄, CO₂R₄,NR₄R₄′, CONR₄R₄′, CH₂OR₄, alkyl or substituted alkyl, alkenyl orsubstituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl orsubstituted cycloalkyl, arylalkyl or substituted arylalkyl, aryl orsubstituted aryl and heteroaryl or substituted heteroaryl; and

W is selected from (CR₆R₆′), C(R₆)OR₃, C(R₆)(NR₄R₄′),

n is an integer of 1 or 2;

including all prodrug esters, pharmaceutically acceptable salts andstereoisomers thereof,

with the following provisos:

(a) when R₅ and R₅′ and/or R₆ and R₆′ form a double bond with CR₇R₇′,when either R₇ or R₇′ is OR₄, R₄ is not hydrogen;

(b) excluding compounds where the following occur simultanously:

R₂ or R₂′ are hydrogen, OR₃, halo, NHCOR₄, NHCO₂R₄, NHCONR₄R₄′ orNHSO₂R₄;

R₅ and R₅′ are hydrogen or form a double bond with oxygen or sulfur;

R₆ and R₆′ are hydrogen, alkyl or substituted alkyl, alkenyl orsubstituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl orsubstituted cycloalkyl, arylalkyl or substituted arylalkyl, aryl orsubstituted aryl, or heteroaryl or substituted heteroaryl, wherein atleast one of R₆ and R₆′ is hydrogen, or R₆ and R₆′ taken together form adouble bond with oxygen (O), sulfur (S) or NR₇;

R₇ is hydrogen, alkyl or substituted alkyl, alkenyl or substitutedalkenyl, alkynyl or substituted alkynyl, cycloalkyl or substitutedcycloalkyl, arylalkyl or substituted arylalkyl, aryl or substitutedaryl, or heteroaryl or substituted heteroaryl; and

G has the following structure:

wherein

R₁₃ is selected from hydrogen (H), cyano (—CN), nitro (—NO₂), halo,heterocyclo, OR₁₄, CO₂R₁₅, CONHR₁₅, COR₁₅, S(O)_(p)R₁₅, SO₂NR₁₅R,₅′,NHCOR₁₅ and NHSO₂R₁₅;

R₁₄ in each functional group is independently selected from hydrogen(H), alkyl or substituted alkyl, CHF₂, CF₃ and COR₁₅;

R₁₅ and R₁₅′ in each functional group are each independently selectedfrom hydrogen(H), alkyl or substituted alkyl, alkenyl or substitutedalkenyl, alkynyl or substituted alkynyl, cycloalkyl or substitutedcycloalkyl, heterocycloalkyl or substituted heterocycloalkyl, arylalkylor substituted arylalkyl, aryl or substituted aryl, heteroaryl orsubstituted heteroaryl and —CN;

A and B are each independently selected from hydrogen (H), halo, cyano(—CN), nitro (—NO₂), alkyl or substituted alkyl and OR₁₄; and

p is an integer from 0 to 2.

In a preferred embodiment, the present invention provides compoundsaccording to formula I, as defined above, wherein G is selected from:

wherein

R₈, R₉, R₁₀ and R₁₁ are each independently selected from hydrogen (H),NO₂, CN, CF₃, OR₄, CO₂R₄, NR₄R₄′, CONR₄R₄′, CH₂OR₄, alkyl or alkyl,alkenyl or substituted alkenyl, alkynyl or substituted alkynyl,cycloalkyl or substituted cycloalkyl, arylalkyl or substitutedarylalkyl, aryl or substituted aryl and heteroaryl or substitutedheteroaryl;

A to F is each independently selected from N or CR₉;

J, K, L, P and Q are each independently selected from NR₁₂, O, S, SO,SO₂ or CR₁₂R₁₂′;

R₁₂ and R₁₂′ in each functional group are each independently selectedfrom a bond or R₁; and

m is an integer of 0 or 1.

In a more preferred embodiment, the present invention provides compoundsaccroding to formula I, as defined above, wherein

R₁ is hydrogen (H) or alkyl;

R₂ or R₂′ is hydroxyl (OH);

R₅ and R₅′ are hydrogen or are taken together form a double bond withoxygen (O) or sulfur (S); and

R₆ and R₆′ are taken together form a double bond with oxygen (O) orsulfur (S).

In a more preferred embodiment, the present invention provides compoundsaccording to formula I, as defined above, wherein R₈ is CN.

In a more preferred embodiment, the present invention provides compoundsaccording to formula I selected from:

In a more preferred embodiment, the present invention provides compoundsaccording to formula I selected from:

In a more preferred embodiment, the present invention provides compoundsaccording to formula I selected from:

In a second embodiment, the present invention provides compoundsaccording to formula Ih

wherein

R₁ is selected from hydrogen (H), alkyl or substituted alkyl, alkenyl orsubstituted alkenyl, arylalkyl or substituted arylalkyl, CO₂R₄, CONR₄R₄′and CH₂OR₄;

R₂ and R₂′ are each independently selected from hydrogen (H), alkyl,substituted alkyl, OR₃, SR₃, halo, NHR₄, NHCOR₄, NHCO₂R₄, NHCONR₄R₄′ andNHSO₂R₄;

and at least one of R₂ and R₂′ is H or alkyl, with the exception that R₂and R₂′ can both be OR₃ when R₃ is not H;

R₃ in each functional group is independently selected from hydrogen (H),alkyl or substituted alkyl, CHF₂, CF₃ and COR₄;

R₄ and R₄′ in each functional group are each independently selected fromhydrogen(H), alkyl or substituted alkyl, alkenyl or substituted alkenyl,alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl,arylalkyl or substituted arylalkyl, aryl or substituted aryl, andheteroaryl or substituted heteroaryl;

X and Y are each independently oxygen (O) or sulfur (S);

G is an aryl, heterocyclo or heteroaryl group, wherein said group ismono- or polycyclic, and which is optionally substituted with one ormore substitutents selected from the group consisting of hydrogen, halo,CN, CF₃, OR₄, CO₂R₄, NR₄R₄′, CONR₄R₄′, CH₂OR₄, alkyl or substitutedalkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl,cycloalkyl or substituted cycloalkyl, arylalkyl or substitutedarylalkyl, aryl or substituted aryl and heteroaryl or substitutedheteroaryl; and

W is selected from (CR₆R₆′), C(R₆)OR₃, C(R₆)(NR₄R₄′),

n is an integer of 1 or 2;

including all prodrug esters, pharmaceutically acceptable salts andstereoisomers thereof,

with the following proviso:

(a) excluding compounds where the following occur simultanously:

R₂ or R₂′ is hydrogen, OR₃, halo, NHCOR₄, NHCO₂R₄, NHCONR₄R₄′ orNHSO₂R₄; and

G has the following structure:

wherein

R₁₃ is selected from hydrogen (H), cyano (—CN), nitro (—NO₂), halo,heterocyclo, OR₁₄, CO₂R₁₅, CONHR₁₅, COR₁₅, S(O)_(p)R₁₅, SO₂NR₁₅R₁₅′,NHCOR₁₅ and NHSO₂R₁₅;

R₁₄ in each functional group is independently selected from (H), alkylor substituted alkyl, CHF₂, CF₃ and COR₁₅;

R₁₅ and R₁₅′ in each functional group are each independently selectedfrom hydrogen(H), alkyl or substituted alkyl, alkenyl or substitutedalkenyl, alkynyl or substituted alkynyl, cycloalkyl or substitutedcycloalkyl, heterocycloalkyl or substituted heterocycloalkyl, arylalkylor substituted arylalkyl, aryl or substituted aryl, heteroaryl orsubstituted heteroaryl and —CN;

A and B are each independently selected from hydrogen (H), halo, cyano(—CN), nitro (—NO₂), alkyl or substituted alkyl and OR₁₄; and

p is an integer from 0 to 2.

In a preferred embodiment, the present invention provides compoundsaccording to formula Ih, as defined above, wherein G is selected from:

wherein

R₈, R₉, R₁₀ and R₁₁ in each functional group are each independentlyselected from hydrogen (H), NO₂, CN, CF₃, OR₄, CO₂R₄, NR₄R₄′, CONR₄R₄′,CH₂OR₄, alkyl or substituted alkyl, alkenyl or substituted alkenyl,alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl,arylalkyl or substituted arylalkyl, aryl or substituted aryl andheteroaryl or substituted heteroaryl;

A to F is each independently selected from N or CR₉;

J, K, L, P and Q are each independently selected from NR₁₂, O, S, SO,SO₂ or CR₁₂R₁₂′;

R₁₂ and R₁₂′ in each functional group are each independently selectedfrom a bond or R₁; and

m is an integer of 0 or 1.

In a more preferred embodiment, the present invention provides compoundsaccording to formula Ih, as defined above, wherein

R₁ is hydrogen (H) or alkyl; and

R₂ or R₂′ is hydroxyl (OH).

In a more preferred embodiment, the present invention provides compoundsaccording to formula Ih, as defined above, wherein R₈ is CN.

In a more preferred embodiment, the present invention providespharmaceutical composition, comprising:

-   -   (a) a compound according to formula I; and    -   (b) at least one pharmaceutically acceptable diluent or carrier.

In a more preferred embodiment, the present invention provides thepharmaceutical compositions defined above, further comprising at leastone additional therapeutic agent selected from other compounds offormula I, parathyroid hormone, bisphosphonates, estrogen, testosterone,progesterone, selective estrogen receptor modulators, growth hormonesecretagogues, growth hormone, progesterone receptor modulators,anti-diabetic agents, anti-hypertensive agents, anti-inflammatoryagents, anti-osteoporosis agents, anti-obesity agents, cardiacglycosides, cholesterol lowering agents, anti-depressants, anti-anxietyagents, anabolic agents, and thyroid mimetics.

In a further preferred embodiment, the present invention provides thepharmaceutical compositions defined above, wherein the additionaltherapeutic agent is selected from the group consisting of growthhormone secretagogues and growth hormone.

In a third embodiment, the present invention provides for a method fortreating or delaying the progression or onset of muscular atrophy,lipodistrophy, long-term critical illness, sarcopenia, frailty orage-related functional decline, reduced muscle strength and function,reduced bone density or growth, the catabolic side effects ofglucocorticoids, chronic fatigue syndrome, bone fracture repair, acutefatigue syndrome and muscle loss following elective surgery, cachexia,chronic catabolic state, eating disorders, side effects of chemotherapy,wasting, depression, nervousness, irritability, stress, growthretardation, reduced cognitive function, male contraception,hypogonadism, Syndrome X, diabetic complications or obesity, whichcomprises administering to a mammalian species in need of treatment atherapeutically effective amount of a pharmaceutical composition asdefined in claim 1.

In a preferred embodiment, the present invention provides the methodsdefined above, further comprising administering, concurrently orsequentially, a therapeutically effective amount of at least oneadditional therapeutic agent selected from the group consisting of othercompounds formula I, parathyroid hormone, bisphosphonates, estrogen,testosterone, progesterone, selective estrogen receptor modulators,growth hormone secretagogues, growth hormone, progesterone receptormodulators, anti-diabetic agents, anti-hypertensive agents,anti-inflammatory agents, anti-osteoporosis agents, anti-obesity agents,cardiac glycosides, cholesterol lowering agents, anti-depressants,anti-anxiety agents, anabolic agents and thyroid mimetics.

In a fourth embodiment, the present invention provides for a process forpreparing a compound of formula Id

which comprises hydrolyzing a compound of formula IVa

under basic conditions to give the compound of formula XIX

which is then carried on to a compound of formula Id with the use of acoupling reagent.

In a fifth embodiment, the present invention provides for a process forpreparing a compound of formula Ie

which comprises optionally protecting the compound of formula IVa, whenR2 is OH, with a protecting group by treatment with a silylating reagentand then reduced with a reducing agent to give a compound of formula XX

which is then derivatized with a leaving group and p-toluenesulfonylchloride and then treated with a base to give the compound of formulaIe.

In a preferred embodiment, the present invention provides for processesdefined above wherein the protecting group is tert-Butyldimethylsilyl;the silylating reagent is tert-Butyldimethylsilyl (chloride); thereducing agent is lithium aluminum hydride or lithium borohydride; theleaving group is Tosyl; the base is potassium tert-butoxide.

In a sixth embodiment, the present invention provides for a process forpreparing a compound of formula XII,

which comprises reacting an aldehyde of formula IX

with an amine of formula XV

in the presence of a reducing agent to give the compound of formula XII.

In a seventh embodiment, the present invention provides for a processfor preparing a compound of formula XIV

which comprises subjecting the compound of formula XII prepared by theprocess of claim 18 to N-deprotection to form a compound of formula XIII

and reacting the compound of formula XIII with phosgene or a phosgeneequivalent in the presence of a base to form the compound of formulaXIV.

The following abbreviations are employed herein:

-   Chiralpak®=Trademark of Chiral Technologies, Inc. Eaton, Pa.-   DBU=1,8-diazabicyclo[5.4.0]undec-7-ene-   AcOH=acetic acid-   DMPU=1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone-   EtOAc=ethyl acetate-   HPLC=high performance liquid chromatography-   MeOH=methanol-   MS or Mass Spec=mass spectrometry-   YMC®=trademark of YMC Co, Ltd., Kyoto, Japan-   CuBr=copper(I) bromide-   CuCN=copper(I) cyanide-   CsF=cesium fluoride-   Et₃N=triethylamine-   DCC=1,3-dicyclohexylcarbodiimide-   DEAD=diethyl azodicarboxylate-   LDA=lithium diisopropylamide-   NMP=1-methyl-2-pyrrolidinone-   KOH=potassium hydroxide-   Pd/C=palladium on activated charcoal-   TFA=trifluoroacetic acid-   THF=tetrahydrofuran-   mp.=melting point-   min=minute(s)-   h=hour(s)-   L=liter-   mL=milliliter-   μL=microliter-   g=gram(s)-   mg=milligram(s)-   mol=moles-   mmol=millimole(s)-   nM=nanomolar-   rt=room temperature

The following definitions apply to the terms as used throughout thisspecification, unless otherwise limited in specific instances.

As used herein, the term “alkyl” denotes branched or unbranchedhydrocarbon chains, preferably having about 1 to about 8 carbons, suchas, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl,tert-butyl, 2-methylpentyl pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl and the like. “Substituted alkyl”includes an alkyl group optionally substituted with one or morefunctional groups which are attached commonly to such chains, such as,hydroxyl, bromo, fluoro, chloro, iodo, mercapto or thio, cyano,alkylthio, heterocyclyl, aryl, heteroaryl, carboxyl, carbalkoyl, alkyl,alkenyl, nitro, amino, alkoxyl, amido, and the like to form alkyl groupssuch as trifluoro methyl, 3-hydroxyhexyl, 2-carboxypropyl,2-fluoroethyl, carboxymethyl, cyanobutyl and the like.

Unless otherwise indicated, the term “cycloalkyl” as employed hereinalone or as part of another group includes saturated or partiallyunsaturated (containing 1 or more double bonds) cyclic hydrocarbongroups containing 1 to 3 rings, including monocyclicalkyl, bicyclicalkyland tricyclicalkyl, containing a total of 3 to 20 carbons forming therings, preferably 3 to 10 carbons, forming the ring and which may befused to 1 or 2 aromatic rings as described for aryl, which includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl, cyclodecyl and cyclododecyl, cyclohexenyl,

“Substituted cycloalkyl” includes a cycloalkyl group optionallysubstituted with 1 or more substituents such as halogen, alkyl, alkoxy,hydroxy, aryl, aryloxy, arylalkyl, cycloalkyl, alkylamido,alkanoylamino, oxo, acyl, arylcarbonylamino, amino, nitro, cyano, thioland/or alkylthio and/or any of the substituents included in thedefinition of “substituted alkyl.”

Unless otherwise indicated, the term “alkenyl” as used herein by itselfor as part of another group refers to straight or branched chainradicals of 2 to 20 carbons, preferably 2 to 12 carbons, and morepreferably 2 to 8 carbons in the normal chain, which include one or moredouble bonds in the normal chain, such as vinyl, 2-propenyl, 3-butenyl,2-butenyl, 4-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 2-heptenyl,3-heptenyl, 4-heptenyl, 3-octenyl, 3-nonenyl, 4-decenyl, 3-undecenyl,4-dodecenyl, 4,8,12-tetradecatrienyl, and the like. “Substitutedalkenyl” includes an alkenyl group optionally substituted with one ormore substituents, such as the substituents included above in thedefinition of “substituted alkyl” and “substituted cycloalkyl.”

Unless otherwise indicated, the term “alkynyl” as used herein by itselfor as part of another group refers to straight or branched chainradicals of 2 to 20 carbons, preferably 2 to 12 carbons and morepreferably 2 to 8 carbons in the normal chain, which include one or moretriple bonds in the normal chain, such as 2-propynyl, 3-butynyl,2-butynyl, 4-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl,3-heptynyl, 4-heptynyl, 3-octynyl, 3-nonynyl, 4-decynyl, 3-undecynyl,4-dodecynyl and the like. “Substituted alkynyl” includes an alkynylgroup optionally substituted with one or more substituents, such as thesubstituents included above in the definition of “substituted alkyl” and“substituted cycloalkyl.”

The terms “arylalkyl”, “arylalkenyl” and “arylalkynyl” as used alone oras part of another group refer to alkyl, alkenyl and alkynyl groups asdescribed above having an aryl substituent. Representative examples ofarylalkyl include, but are not limited to, benzyl, 2-phenylethyl,3-phenylpropyl, phenethyl, benzhydryl and naphthylmethyl and the like.“Substituted arylalkyl” includes arylalkyl groups wherein the arylportion is optionally substituted with one or more substituents, such asthe substituents included above in the definition of “substituted alkyl”and “substituted cycloalkyl.”

The term “halogen” or “halo” as used herein alone or as part of anothergroup refers to chlorine, bromine, fluorine, and iodine.

Unless otherwise indicated, the term “aryl” or “Ar” as employed hereinalone or as part of another group refers to monocyclic and polycyclicaromatic groups containing 6 to 10 carbons in the ring portion (such asphenyl or naphthyl including 1-naphthyl and 2-naphthyl) and mayoptionally include one to three additional rings fused to a carbocyclicring or a heterocyclic ring (such as aryl, cycloalkyl, heteroaryl orcycloheteroalkyl rings), for example

“Substituted aryl” includes an aryl group optionally substituted withone or more functional groups, such as halo, haloalkyl, alkyl,haloalkyl, alkoxy, haloalkoxy, alkenyl, trifluoromethyl,trifluoromethoxy, alkynyl, cycloalkyl-alkyl, cycloheteroalkyl,cycloheteroalkylalkyl, aryl, heteroaryl, arylalkyl, aryloxy,aryloxyalkyl, arylalkoxy, alkoxycarbonyl, arylcarbonyl, arylalkenyl,aminocarbonylaryl, arylthio, arylsulfinyl, arylazo, heteroarylalkyl,heteroarylalkenyl, heteroarylheteroaryl, heteroaryloxy, hydroxy, nitro,cyano, amino, substituted amino wherein the amino includes 1 or 2substituents (which are alkyl, aryl or any of the other aryl compoundsmentioned in the definitions), thiol, alkylthio, arylthio,heteroarylthio, arylthioalkyl, alkoxyarylthio, alkylcarbonyl,arylcarbonyl, alkylaminocarbonyl, arylaminocarbonyl, alkoxycarbonyl,aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino,arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfonylamino orarylsulfonaminocarbonyl and/or any of the alkyl substituents set outherein.

Unless otherwise indicated, the term “heteroaryl” as used herein aloneor as part of another group refers to a 5- or 7-membered aromatic ringwhich includes 1, 2, 3 or 4 hetero atoms such as nitrogen, oxygen orsulfur and such rings fused to an aryl, cycloalkyl, heteroaryl orheterocycloalkyl ring (e.g. benzothiophenyl, indolyl), and includespossible N-oxides. “Substituted heteroaryl” includes a heteroaryl groupoptionally substituted with 1 to 4 substituents, such as thesubstituents included above in the definition of “substituted alkyl” and“substituted cycloalkyl.” Examples of heteroaryl groups include thefollowing:

and the like.

The term “heterocyclo”, heterocycle or heterocyclic ring, as usedherein, represents an unsubstituted or substituted stable 5- to7-membered monocyclic ring system which may be saturated or unsaturated,and which consists of carbon atoms and from one to four heteroatomsselected from N, O or S, and wherein the nitrogen and sulfur heteroatomsmay optionally be oxidized, and the nitrogen heteroatom may optionallybe quaternized. The heterocyclic ring may be attached at any heteroatomor carbon atom which results in the creation of a stable structure.Examples of such heterocyclic groups include, but is not limited to,piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidinyl,oxopyrrolidinyl, oxoazepinyl, azepinyl, pyrrolyl, pyrrolidinyl, furanyl,thienyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl,imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl,oxazolidinyl, isooxazolyl, isoxazolidinyl, morpholinyl, thiazolyl,thiazolidinyl, isothiazolyl, thiadiazolyl, tetrahydropyranyl,thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, andoxadiazolyl.

The compounds of formula I can be present as salts, which are alsowithin the scope of this invention. Pharmaceutically acceptable (i.e.,non-toxic, physiologically acceptable) salts are preferred. If thecompounds of formula I have, for example, at least one basic center,they can form acid addition salts. These are formed, for example, withstrong inorganic acids, such as mineral acids, for example sulfuricacid, phosphoric acid or a hydrohalic acid, with strong organiccarboxylic acids, such as alkanecarboxylic acids of 1 to 4 carbon atomswhich are unsubstituted or substituted, for example, by halogen, forexample acetic acid, such as saturated or unsaturated dicarboxylicacids, for example oxalic, malonic, succinic, maleic, fumaric, phthalicor terephthalic acid, such as hydroxycarboxylic acids, for exampleascorbic, glycolic, lactic, malic, tartaric or citric acid, such asamino acids, (for example aspartic or glutamic acid or lysine orarginine), or benzoic acid, or with organic sulfonic acids, such as(C₁-C₄) alkyl or arylsulfonic acids which are unsubstituted orsubstituted, for example by halogen, for example methyl- orp-toluene-sulfonic acid. Corresponding acid addition salts can also beformed having, if desired, an additionally present basic center. Thecompounds of formula I having at least one acid group (for example COOH)can also form salts with bases. Suitable salts with bases are, forexample, metal salts, such as alkali metal or alkaline earth metalsalts, for example sodium, potassium or magnesium salts, or salts withammonia or an organic amine, such as morpholine, thiomorpholine,piperidine, pyrrolidine, a mono, di or tri-lower alkylamine, for exampleethyl, tert-butyl, diethyl, diisopropyl, triethyl, tributyl ordimethyl-propylamine, or a mono, di or trihydroxy lower alkylamine, forexample mono, di or triethanolamine. Corresponding internal salts mayfurthermore be formed. Salts which are unsuitable for pharmaceuticaluses but which can be employed, for example, for the isolation orpurification of free compounds of formula I or their pharmaceuticallyacceptable salts, are also included.

Preferred salts of the compounds of formula I which contain a basicgroup include monohydrochloride, hydrogensulfate, methanesulfonate,phosphate or nitrate.

Preferred salts of the compounds of formula I which contain an acidgroup include sodium, potassium and magnesium salts and pharmaceuticallyacceptable organic amines.

The term “modulator” refers to a chemical compound with capacity toeither enhance (e.g., “agonist” activity) or inhibit (e.g., “antagonist”activity) a functional property of biological activity or process (e.g.,enzyme activity or receptor binding); such enhancement or inhibition maybe contingent on the occurrence of a specific event, such as activationof a signal transduction pathway, and/or may be manifest only inparticular cell types.

The term “prodrug esters” as employed herein includes esters andcarbonates formed by reacting one or more hydroxyls of compounds offormula I with alkyl, alkoxy, or aryl substituted acylating agentsemploying procedures known to those skilled in the art to generateacetates, pivalates, methylcarbonates, benzoates and the like.

Any compound that can be converted in vivo to provide the bioactiveagent (i.e., the compound of formula I) is a prodrug within the scopeand spirit of the invention.

Various forms of prodrugs are well known in the art. A comprehensivedescription of prodrugs and prodrug derivatives are described in:

-   -   a.) The Practice of Medicinal Chemistry, Camille G. Wermuth et        al., Ch 31, (Academic Press, 1996);    -   b.) Design of Prodrugs, edited by H. Bundgaard, (Elsevier,        1985);    -   c.) A Textbook of Drug Design and Development, P.        Krogsgaard-Larson and H. Bundgaard, eds. Ch 5, pgs 113-191        (Harwood Academic Publishers, 1991).        Said references are incorporated herein by reference.

An administration of a therapeutic agent of the invention includesadministration of a therapeutically effective amount of the agent of theinvention. The term “therapeutically effective amount” as used hereinrefers to an amount of a therapeutic agent to treat or prevent acondition treatable by administration of a composition of the invention.That amount is the amount sufficient to exhibit a detectable therapeuticor preventative or ameliorative effect. The effect may include, forexample, treatment or prevention of the conditions listed herein. Theprecise effective amount for a subject will depend upon the subject'ssize and health, the nature and extent of the condition being treated,recommendations of the treating physician, and the therapeutics orcombination of therapeutics selected for administration. Thus, it is notuseful to specify an exact effective amount in advance.

All stereoisomers of the compounds of the instant invention arecontemplated, either in admixture or in pure or substantially pure form.The compounds of the present invention can have asymmetric centers atany of the carbon atoms including any one of the R substituents.Consequently, compounds of formula I can exist in enantiomeric ordiastereomeric forms or in mixtures thereof. The processes forpreparation can utilize racemates, enantiomers or diastereomers asstarting materials. When diastereomeric or enantiomeric products areprepared, they can be separated by conventional methods for example,chromatographic, chiral HPLC or fractional crystallization.

The compounds of formula I of the invention can be prepared as shown inthe following reaction schemes and description thereof, as well asrelevant published literature procedures that may be used by one skilledin the art. Exemplary reagents and procedures for these reactions appearhereinafter and in the working Examples.

As illustrated in Scheme I, compounds of formula Ia can be prepared fromsuitably protected intermediates of formula II. Intermediates of formulaII can be obtained commercially, can be prepared by methods known in theliterature or can be readily prepared by one skilled in the art.Treatment of II with an intermediate of formula III yields anintermediate of formula of IV. The intermediates of formula III can beobtained, for example, from commercially available isocyanates andthioisocyanates and or can be readily prepared by one skilled in theart. The intermediate of formula IV can be treated with a base, such asDBU, to yield a compound of formula Ia. Compounds of formula Iarepresent compounds of formula I wherein R₁ is H, R₅ and R₅′ are takentogether to form a double bond with O or S and R₆ and R₆′ are takentogether to form a double bond with O or S. As illustrated in Scheme Ia,compounds of formula Id and Ie can be prepared from suitably protectedintermediates of formula II by reacting with a compound of formula IIIato form an intermediate of formula IVa. An intermediate of the formulaIVa can be hydrolysed under basic conditions to give an intermediate ofthe formula XIX and then carried on to a compound of the formula Id withthe use of a suitable coupling reagents such as, for example DCC.Alternatively, an intermediate of the formula IVa can be optionallyprotected (where R₂=OH) with a suitable protecting group such as TBS bytreatment with a silylating reagent such as TBDMSCl, and then reducedwith a suitable reducing agent such as, for example LAH or LiBH₄ to givean intermediate of the formula XX. An intermediate of the formula XX canthen be derivatized on the primary hydroxyl functionality with asuitable leaving group such as Tosyl, with, for example,p-toluenesulfonyl chloride, followed by base treatment such as withpotassium tert-butoxide to give a compound of the formula Ie.

As illustrated in Scheme II, a compound of formula Ib, wherein R₁ is H,can be converted to a compound of formula Ic wherein R₁ is a functionalgroup other than H, as defined herein, by treatment with a base such asLDA and an alkyl halide, such as iodomethane, preferably in a solventsuch as THF at low temperatures (e.g., −78° C.). Compounds of formula Icrepresent compounds of formula I wherein R₁ is a functional group otherthan H and R₅ and R₅′ are taken together to form a double bond with O.Optionally, subsequent reaction of compounds of formula Ic with aLawesson's Reagent will convert Y from oxygen (O) to sulfur (S).

As illustrated in Scheme III, a compound of formula Ic can be convertedby treatment with a reducing agent, such as LiEt₃BH, preferably in asolvent such as THF at low temperatures (<−40° C.) to give anintermediate V. Intermediate V is subsequently treated further withEt₃SiH in the presence of boron trifluoride diethyl etherate in ahalogenated solvent such as 1,2-dichloroethane at low temperatures (<0°C.) to yield a compound of formula VI. Compounds of formula VI representcompounds of formula I wherein R₅ and R₅′ are hydrogen. A compound offormula VI can be oxidized to a compound of formula VIa using standardconditions of known oxidation methods, such as, for example, Swern orDess-Martin.

Scheme IV describes a method for preparing compounds of formula XIV fromN-protected amino acids of formula VII, which can be obtainedcommercially or can be prepared by methods known in the literature orcan be readily prepared by one skilled in the art. An intermediate offormula VII is treated with a reducing agent, such as borane, to form analcohol intermediate VIII, which can be oxidized to an aldehydeintermediate IX. Similarly, an intermediate of formula VII can becoupled to N,O-dimethylhydroxylamine to form amide X, which can betreated with a Grignard reagent or an organolithium reagent to form analkylketone XI. The aldehyde intermediate IX or an alkylketone XI, canbe reacted with an amine of formula XV in the presence of a reducingagent, such as sodium triacetoxyborohydride to give an intermediate offormula XII. Removal of N-protecting group (L) can be achieved bymethods known in the literature or by one skilled in the art to providean intermediate of formula XIII. The intermediate of formula XIII can betreated with phosgene or phosgene equivalent in the presence of a base,such as triethylamine, to provide a compound of formula XIV. Compoundsof formula XIV represent compounds of formula I wherein R₆ and R₆′ aretaken together to form a double bond with oxygen and R₅ is hydrogen andR₅′ is as defined herein. In the alternative, Scheme IV may be utilizedto provide compounds of formula I wherein R₅′ is hydrogen and R₅ is asdefined herein. Optionally, subsequent reaction of compounds of formulaXIV with a Lawesson's Reagent will provide compounds of formula Iwherein R₆ and R₆′ are taken together to form a double bond with sulfur(S).

As illustrated in Scheme V, an intermediate of formula VII as describedin Scheme IV can be coupled to an amine XV using a coupling reagent,such as those described in “The Practice of Peptide Synthesis”(Spring-Verlag, 2^(nd) Ed., Bodanszy, Miklos, 1993), to yield an amideintermediate of formula XVI. Removal of N-protecting group can beachieved by methods known in the literature or by one skilled in the artto provide an intermediate of formula XVII. The intermediate of formulaXVII is treated with an aldehyde (R₆CHO) in suitable solvent (such asethanol, methanol, THF or CH₂Cl₂), with or without the presence of abase, such as K₂CO₃, NaOH or DBU, or a weak acid, such as HOAc, to givea compound of formula XVIII. Aldehydes of formula R₆CHO can be obtainedfrom commercial sources, can be prepared by methods known in theliterature or readily prepared by one skilled in the art. Compounds offormula XVIII represent compounds of formula I wherein R₅ and R₅′ aretaken together to form a double bond with oxygen (O) and R₆′ is hydrogenand R₆ is as defined herein. In the alternative, Scheme V may beutilized to provide compounds of formula I wherein R₆ is hydrogen andR₆′ is as defined herein. Optionally, subsequent reaction of compoundsof formula XVIII with a Lawesson's Reagent will provide compounds offormula I wherein R₅ and R₅′ are taken together to form a double bondwith sulfur (S).

Scheme VI describes a method to prepare isocyanates of general formulaIII wherein intermediates XV are treated with phosgene or a phosgenelike reagent in the presence of an inorganic base such as sodiumbicarbonate, or a organic base such as diisopropylethylamine in asolvent such as dichloromethane to afford an isocyanate of formula III.

For example, Scheme VIa describes a method for preparing isocyanates ofgeneral formula IIIa. Substituted aryl or heteroaryl amines of formulaXV are treated with phosgene or a phosgene like reagent in the presenceof an inorganic base such as sodium bicarbonate, or a organic base suchas diisopropylethylamine in a solvent such as dichloromethane to affordan isocyanate of formula IIIa. Substituted aryl or heteroaryl amines asdescribed above can be obtained commercially or can be prepared bymethods known in the literature or by one skilled in the art.

Scheme VII describes a method to prepare aryl and heteroaryl halides ofgeneral formula XXI and XXII wherein intermediates XV are treated withtert-butylnitrite or an equivalent reagent in the presence of a halidesource such as CuI or CuBr to afford an aryl or heteroaryl halide offormula XXI or XXII which are precursors suitable for the formation ofthe intermediates of formula XII.

Scheme VIII describes additional methods for preparing compounds offormula XIV from N-protected aldehydes of formula IX. An intermediate offormula IX is treated with an organometalic reagent such as R_(5′)MgX (Xis typically a halide), R_(5′)Li or alternate alkylating agents such asTMS-CF₃ with CsF or equivalent reagents to form an alcohol intermediateXXIII, which can be oxidized to ketone intermediate XI. The ketoneintermediate XI can be reacted with an amine such as hydroxylamine or anequivalent reagent to form an imine of formula XXIV. Intermediates offormula XXIV in the presence of a reducing agents, such as hydrogen inthe presence of palladium or nickel catalysts, give an amineintermediate of formula XXV. Coupling of intermediates of formula XXVwith halides of formula XXI or XXII to give intermediates of formula XIIcan be achieved by methods known in the literature or by one skilled inthe art. The intermediate of formula XII can be treated as described inScheme IV to provide a compound of formula XIV. Alternately,intermediates of formula XXIII can be converted to intermediates offormula XXV through a three step process involving activation of thealcohol with a reagent such as mesyl or tosyl chloride in the presenceof an organic or inorganic base followed by treatment with an azidesource such as sodium azide and finally reduction of the azide to theamine to provide intermediates of formula XXV.

Scheme IX describes additional methods for preparing compounds offormula XIV from N-protected alcohols of formula XXIII. Removal ofN-protecting group (L) can be achieved by methods known in theliterature or by one skilled in the art to provide an intermediate offormula XXVI. An intermediate of formula XXVI is treated with anisocyanate of formula III to give an intermediate of formula XXVII.Treatment of intermediates of formula XXVII with an activating agentsuch as mesyl chloride or tosyl chloride in the presence of a base suchas potassium tert-butoxide or an equivalent reagent can provide acompound of formula XIV.

A hydroxyl group on the bicyclic core present in some compounds of thepresent genus can be replaced with alternative functionality, includinghalogens and amine-based groups according to the sequences shown inScheme X. For example, a compound of formula XXVIII can be treated witha suitable fluorinating reagent, such as, for example, DAST, to providecompounds of formula XXIX. Compounds of formula XXVIII can alternativelybe derivatised with a suitable functionality to generate a leavinggroup, such as, for example, conversion to a mesyl group under standardconditions to afford a compound of the formula XXX, which can thenundergo a substitution reaction with a suitable nitrogen basednucleophile, such as, for example, azide anion to afford an intermediateof the formula XXXI. Intermediate azides such as those of the formulaXXXI can then be reduced to the corresponding amine with a suitablereducing agent, such as, for example, PtO₂, to yield a primary aminesuch as found in compounds of the formula XXXII. Compounds of theformula XXXII can readily undergo N-acylations or N-alkylations understandard conditions to afford compounds of the formulas XXXIII andXXXIV, respectively.

In addition, a compound of formula XXVIII can be treated with a suitableoxidizing agent, such as, for example, TPAP with NMO, to provideintermediates of formula XXXV. Treatment of intermediates of formulaXXXV with an amine, such as hydroxyl amine or an equivalent reagent, cangive a compound of formula XXXVI.

Furthermore, alcohol diastereomers of compounds such as XXVIII can beobtained by inversion of the alcohol by methods such as the Mitsunobureaction followed by hydrolysis or oxidation followed by reduction orother methods familiar to those trained in the art.

Use and UtilityA. Utilities

The compounds of the present invention modulate the function of thenuclear hormone receptors, particularly the androgen receptor, andinclude compounds which are, for example, selective agonists, partialagonists, antagonists or partial antagonists of the androgen receptor(AR). Thus, the present compounds are useful in the treatment ofAR-associated conditions. An “AR-associated condition,” as used herein,denotes a condition or disorder which can be treated by modulating thefunction or activity of an AR in a subject, wherein treatment comprisesprevention, partial alleviation or cure of the condition or disorder.Modulation may occur locally, for example, within certain tissues of thesubject, or more extensively throughout a subject being treated for sucha condition or disorder.

The compounds of the present invention can be administered to animals,preferably humans, for the treatment of a variety of conditions anddisorders, including, but not limited to maintenance of muscle strengthand function (e.g., in the elderly); reversal or prevention of frailtyor age-related functional decline (“ARFD”) in the elderly (e.g.,sarcopenia); treatment of catabolic side effects of glucocorticoids;prevention and/or treatment of reduced bone mass, density or growth(e.g., osteoporosis and osteopenia); treatment of chronic fatiguesyndrome (CFS); chronic myalgia; treatment of acute fatigue syndrome andmuscle loss following elective surgery (e.g., post-surgicalrehabilitation); accelerating of wound healing; accelerating bonefracture repair (such as accelerating the recovery of hip fracturepatients); accelerating healing of complicated fractures, e.g.distraction osteogenesis; in joint replacement; prevention ofpost-surgical adhesion formation; acceleration of tooth repair orgrowth; maintenance of sensory function (e.g., hearing, sight,olefaction and taste); treatment of periodontal disease; treatment ofwasting secondary to fractures and wasting in connection with chronicobstructive pulmonary disease (COPD), chronic liver disease, AIDS,weightlessness, cancer cachexia, burn and trauma recovery, chroniccatabolic state (e.g., coma), eating disorders (e.g., anorexia) andchemotherapy; treatment of cardiomyopathy; treatment ofthrombocytopenia; treatment of growth retardation in connection withCrohn's disease; treatment of short bowel syndrome; treatment ofirritable bowel syndrome; treatment of inflammatory bowel disease;treatment of Crohn's disease and ulcerative colits; treatment ofcomplications associated with transplantation; treatment ofphysiological short stature including growth hormone deficient childrenand short stature associated with chronic illness; treatment of obesityand growth retardation associated with obesity; treatment of anorexia(e.g., associated with cachexia or aging); treatment of hypercortisolismand Cushing's syndrome; Paget's disease; treatment of osteoarthritis;induction of pulsatile growth hormone release; treatment ofosteochondrodysplasias; treatment of depression, nervousness,irritability and stress; treatment of reduced mental energy and lowself-esteem (e.g., motivation/assertiveness); improvement of cognitivefunction (e.g., the treatment of dementia, including Alzheimer's diseaseand short term memory loss); treatment of catabolism in connection withpulmonary dysfunction and ventilator dependency; treatment of cardiacdysfunction (e.g., associated with valvular disease, myocardialinfarction, cardiac hypertrophy or congestive heart failure); loweringblood pressure; protection against ventricular dysfunction or preventionof reperfusion events; treatment of adults in chronic dialysis; reversalor slowing of the catabolic state of aging; attenuation or reversal ofprotein catabolic responses following trauma (e.g., reversal of thecatabolic state associated with surgery, congestive heart failure,cardiac myopathy, burns, cancer, COPD etc.); reducing cachexia andprotein loss due to chronic illness such as cancer or AIDS; treatment ofhyperinsulinemia including nesidioblastosis; treatment ofimmunosuppressed patients; treatment of wasting in connection withmultiple sclerosis or other neurodegenerative disorders; promotion ofmyelin repair; maintenance of skin thickness; treatment of metabolichomeostasis and renal homeostasis (e.g., in the frail elderly);stimulation of osteoblasts, bone remodeling and cartilage growth;regulation of food intake; treatment of insulin resistance, includingNIDDM, in mammals (e.g., humans); treatment of insulin resistance in theheart; improvement of sleep quality and correction of the relativehyposomatotropism of senescence due to high increase in REM sleep and adecrease in REM latency; treatment of hypothermia; treatment ofcongestive heart failure; treatment of lipodystrophy (e.g., in patientstaking HIV or AIDS therapies such as protease inhibitors); treatment ofmuscular atrophy (e.g., due to physical inactivity, bed rest or reducedweight-bearing conditions); treatment of musculoskeletal impairment(e.g., in the elderly); improvement of the overall pulmonary function;treatment of sleep disorders; and the treatment of the catabolic stateof prolonged critical illness; treatment of hirsutism, acne, seborrhea,androgenic alopecia, anemia, hyperpilosity, benign prostate hypertrophy,adenomas and neoplasies of the prostate (e.g., advanced metastaticprostate cancer) and malignant tumor cells containing the androgenreceptor, such as is the case for breast, brain, skin, ovarian, bladder,lymphatic, liver and kidney cancers; cancers of the skin, pancreas,endometrium, lung and colon; osteosarcoma; hypercalcemia of malignancy;metastatic bone disease; treatment of spermatogenesis, endometriosis andpolycystic ovary syndrome; conteracting preeclampsia, eclampsia ofpregnancy and preterm labor; treatment of premenstural syndrome;treatment of vaginal dryness; age related decreased testosterone levelsin men, male menopause, hypogonadism, male hormone replacement, male andfemale sexual dysfunction (e.g., erectile dysfunction, decreased sexdrive, sexual well-being, decreased libido), urinary incontinence, maleand female contraception, hair loss, Reaven's Syndrome and theenhancement of bone and muscle performance/strength. The term treatmentis also intended to include prophylactic treatment.

In addition, the conditions, diseases, and maladies collectivelyreferenced to as “Syndrome X” or Metabolic Syndrome as detailed inJohannsson J. Clin. Endocrinol. Metab., 82, 727-34 (1997), may betreated employing the compounds of the invention.

B. Combinations

The present invention includes within its scope pharmaceuticalcompositions comprising, as an active ingredient, a therapeuticallyeffective amount of at least one of the compounds of formula I, alone orin combination with a pharmaceutical carrier or diluent. Optionally,compounds of the present invention can be used alone, in combinationwith other compounds of the invention, or in combination with one ormore other therapeutic agent(s), e.g., an antibiotic or otherpharmaceutically active material.

The compounds of the present invention may be combined with growthpromoting agents, such as, but not limited to, TRH, diethylstilbesterol,theophylline, enkephalins, E series prostaglandins, compounds disclosedin U.S. Pat. No. 3,239,345, e.g., zeranol, and compounds disclosed inU.S. Pat. No. 4,036,979, e.g., sulbenox or peptides disclosed in U.S.Pat. No. 4,411,890.

The compounds of the invention may also be used in combination withgrowth hormone secretagogues such as GHRP-6, GHRP-1 (as described inU.S. Pat. No. 4,411,890 and publications WO 89/07110 and WO 89/07111),GHRP-2 (as described in WO 93/04081), NN703 (Novo Nordisk), LY444711(Lilly), MK-677 (Merck), CP424391 (Pfizer) and B-HT920, or with growthhormone releasing factor and its analogs or growth hormone and itsanalogs or somatomedins including IGF-1 and IGF-2, or withalpha-adrenergic agonists, such as clonidine or serotinin 5-HT_(D)agonists, such as sumatriptan, or agents which inhibit somatostatin orits release, such as physostigmine and pyridostigmine. A still furtheruse of the disclosed compounds of the invention is in combination withparathyroid hormone, PTH(1-34) or bisphosphonates, such as MK-217(alendronate).

A still further use of the compounds of the invention is in combinationwith estrogen, testosterone, a selective estrogen receptor modulator,such as tamoxifen or raloxifene, or other androgen receptor modulators,such as those disclosed in Edwards, J. P. et. al., Bio. Med. Chem. Let.,9, 1003-1008 (1999) and Hamann, L. G. et. al., J. Med. Chem., 42,210-212 (1999).

A further use of the compounds of this invention is in combination withprogesterone receptor agonists (“PRA”), such as levonorgestrel,medroxyprogesterone acetate (MPA).

The compounds of the present invention may be employed alone or incombination with each other and/or other modulators of nuclear hormonereceptors or other suitable therapeutic agents useful in the treatmentof the aforementioned disorders including: anti-diabetic agents;anti-osteoporosis agents; anti-obesity agents; anti-inflammatory agents;anti-anxiety agents; anti-depressants; anti-hypertensive agents;anti-platelet agents; anti-thrombotic and thrombolytic agents; cardiacglycosides; cholesterol/lipid lowering agents; mineralocorticoidreceptor antagonists; phospodiesterase inhibitors; protein tyrosinekinase inhibitors; thyroid mimetics (including thyroid receptoragonists); anabolic agents; HIV or AIDS therapies; therapies useful inthe treatment of Alzheimer's disease and other cognitive disorders;therapies useful in the treatment of sleeping disorders;anti-proliferative agents; and anti-tumor agents.

Examples of suitable anti-diabetic agents for use in combination withthe compounds of the present invention include biguanides (e.g.,metformin), glucosidase inhibitors (e.g., acarbose), insulins (includinginsulin secretagogues or insulin sensitizers), meglitinides (e.g.,repaglinide), sulfonylureas (e.g., glimepiride, glyburide andglipizide), biguanide/glyburide combinations (e.g., Glucovance®),thiazolidinediones (e.g., troglitazone, rosiglitazone and pioglitazone),PPAR-alpha agonists, PPAR-gamma agonists, PPAR alpha/gamma dualagonists, SGLT2 inhibitors, glycogen phosphorylase inhibitors,inhibitors of fatty acid binding protein (aP2) such as those disclosedin U.S. Ser. No. 09/519,079 filed Mar. 6, 2000, glucagon-like peptide-1(GLP-1), and dipeptidyl peptidase IV (DPP4) inhibitors such as thosedisclosed in WO 0168603.

Examples of suitable anti-osteoporosis agents for use in combinationwith the compounds of the present invention include alendronate,risedronate, PTH, PTH fragment, raloxifene, calcitonins, steroidal ornon-steroidal progesterone receptor agonists, RANK ligand antagonists,calcium sensing receptor antagonists, TRAP inhibitors, selectiveestrogen receptor modulators (SERM's), estrogen and AP-1 inhibitors.

Examples of suitable anti-obesity agents for use in combination with thecompounds of the present invention include aP2 inhibitors, such as thosedisclosed in U.S. Ser. No. 09/519,079 filed Mar. 6, 2000, PPAR gammaantagonists, PPAR delta agonists, beta 3 adrenergic agonists, such asAJ9677 (Takeda/Dainippon), L750355 (Merck), or CP331648 (Pfizer) orother known beta 3 agonists as disclosed in U.S. Pat. Nos. 5,541,204,5,770,615, 5,491,134, 5,776,983 and 5,488,064, a lipase inhibitor, suchas orlistat or ATL-962 (Alizyme), a serotonin (and dopamine) reuptakeinhibitor, such as sibutramine, topiramate (Johnson & Johnson) oraxokine (Regeneron), a thyroid receptor beta drug, such as a thyroidreceptor ligand as disclosed in WO 97/21993 (U. Cal SF), WO 99/00353(KaroBio) and GB98/284425 (KaroBio), and/or an anorectic agent, such asdexamphetamine, phentermine, phenylpropanolamine or mazindol.

Examples of suitable anti-inflammatory agents for use in combinationwith the compounds of the present invention include prednisone,dexamethasone, Enbrel®, cyclooxygenase inhibitors (i.e., COX-1 and/orCOX-2 inhibitors such as NSAIDs, aspirin, indomethacin, ibuprofen,piroxicam, Naproxen®, Celebrex®, Vioxx®), CTLA4-Ig agonists/antagonists,CD40 ligand antagonists, IMPDH inhibitors, such as mycophenolate(CellCept®), integrin antagonists, alpha-4 beta-7 integrin antagonists,cell adhesion inhibitors, interferon gamma antagonists, ICAM-1, tumornecrosis factor (TNF) antagonists (e.g., infliximab, OR₁₃₈₄),prostaglandin synthesis inhibitors, budesonide, clofazimine, CNI-1493,CD4 antagonists (e.g., priliximab), p38 mitogen-activated protein kinaseinhibitors, protein tyrosine kinase (PTK) inhibitors, IKK inhibitors,and therapies for the treatment of irritable bowel syndrome (e.g.,Zelmac® and Maxi-K® openers such as those disclosed in U.S. Pat. No.6,184,231 B1).

Examples of suitable anti-anxiety agents for use in combination with thecompounds of the present invention include diazepam, lorazepam,buspirone, oxazepam, and hydroxyzine pamoate.

Examples of suitable anti-depressants for use in combination with thecompounds of the present invention include citalopram, fluoxetine,nefazodone, sertraline, and paroxetine.

Examples of suitable anti-hypertensive agents for use in combinationwith the compounds of the present invention include beta adrenergicblockers, calcium channel blockers (L-type and T-type; e.g. diltiazem,verapamil, nifedipine, amlodipine and mybefradil), diuretics (e.g.,chlorothiazide, hydrochlorothiazide, flumethiazide, hydroflumethiazide,bendroflumethiazide, methylchlorothiazide, trichloromethiazide,polythiazide, benzthiazide, ethacrynic acid tricrynafen, chlorthalidone,furosemide, musolimnine, bumetanide, triamtrenene, amiloride,spironolactone), renin inhibitors, ACE inhibitors (e.g., captopril,zofenopril, fosinopril, enalapril, ceranopril, cilazopril, delapril,pentopril, quinapril, ramipril, lisinopril), AT-1 receptor antagonists(e.g., losartan, irbesartan, valsartan), ET receptor antagonists (e.g.,sitaxsentan, atrsentan and compounds disclosed in U.S. Pat. Nos.5,612,359 and 6,043,265), Dual ET/AII antagonist (e.g., compoundsdisclosed in WO 00/01389), neutral endopeptidase (NEP) inhibitors,vasopepsidase inhibitors (dual NEP-ACE inhibitors) (e.g., omapatrilatand gemopatrilat), and nitrates.

Examples of suitable anti-platelet agents for use in combination withthe compounds of the present invention include GPIIb/IIIa blockers(e.g., abciximab, eptifibatide, tirofiban), P2Y12 antagonists (e.g.,clopidogrel, ticlopidine, CS-747), thromboxane receptor antagonists(e.g., ifetroban), aspirin, and PDE-III inhibitors (e.g., dipyridamole)with or without aspirin.

Examples of suitable cardiac glycosides for use in combination with thecompounds of the present invention include digitalis and ouabain.

Examples of suitable cholesterol/lipid lowering agents for use incombination with the compounds of the present invention include HMG-CoAreductase inhibitors (e.g., pravastatin, lovastatin, atorvastatin,simvastatin, NK-104 (a.k.a. itavastatin, or nisvastatin or nisbastatin)and ZD-4522 (a.k.a. rosuvastatin, or atavastatin or visastatin)),squalene synthetase inhibitors, fibrates, bile acid sequestrants, ACATinhibitors, MTP inhibitors, lipooxygenase inhibitors, cholesterolabsorption inhibitors, and cholesterol ester transfer protein inhibitors(e.g., CP-529414).

Examples of suitable mineralocorticoid receptor antagonists for use incombination with the compounds of the present invention includespironolactone and eplerinone.

Examples of suitable phospodiesterase (PDE) inhibitors for use incombination with the compounds of the present invention include PDE-3inhibitors such as cilostazol, and phosphodiesterase-5 inhibitors (PDE-5inhibitors) such as sildenafil.

Examples of suitable thyroid mimetics for use in combination with thecompounds of the present invention include thyrotropin, polythyroid,KB-130015, and dronedarone.

Examples of suitable anabolic agents for use in combination with thecompounds of the present invention include testosterone, TRHdiethylstilbesterol, estrogens, β-agonists, theophylline, anabolicsteroids, dehydroepiandrosterone, enkephalins, E-series prostagladins,retinoic acid and compounds as disclosed in U.S. Pat. No. 3,239,345,e.g., Zeranol®; U.S. Pat. No. 4,036,979, e.g., Sulbenox® or peptides asdisclosed in U.S. Pat. No. 4,411,890.

Examples of suitable HIV or AIDS therapies for use in combination withthe compounds of the present invention include indinavir sulfate,saquinavir, saquinavir mesylate, ritonavir, lamivudine, zidovudine,lamivudine/zidovudine combinations, zalcitabine, didanosine, stavudine,and megestrol acetate.

Examples of suitable therapies for treatment of Alzheimer's disease andcognitive disorders for use in combination with the compounds of thepresent invention include donepezil, tacrine, revastigmine, 5HT6, gammasecretase inhibitors, beta secretase inhibitors, SK channel blockers,Maxi-K blockers, and KCNQs blockers.

Examples of suitable therapies for treatment of sleeping disorders foruse in combination with the compounds of the present invention includemelatonin analogs, melatonin receptor antagonists, ML1B agonists, andGABA/NMDA receptor antagonists.

Examples of suitable anti-proliferative agents for use in combinationwith the compounds of the present invention include cyclosporin A,paclitaxel, FK-506, and adriamycin.

Examples of suitable anti-tumor agents for use in combination with thecompounds of the present invention include paclitaxel, adriamycin,epothilones, cisplatin and carboplatin.

Compounds of the present invention may further be used in combinationwith nutritional supplements such as those described in U.S. Pat. No.5,179,080, especially in combination with whey protein or casein, aminoacids (such as leucine, branched amino acids and hydroxymethylbutyrate),triglycerides, vitamins (e.g., A, B6, B12, folate, C, D and E), minerals(e.g., selenium, magnesium, zinc, chromium, calcium and potassium),carnitine, lipoic acid, creatinine, B-hyroxy-B-methylbutyriate (Juven)and coenzyme Q-10.

In addition, compounds of the present invention may be used incombination with therapeutic agents used in the treatment of sexualdysfunction, including but not limited to PDE-5 inhibitors, such assildenafil or IC-351.

Compounds of the present invention may further be used in combinationwith antiresorptive agents, hormone replacement therapies, vitamin Danalogues, elemental calcium and calcium supplements, cathepsin Kinhibitors, MMP inhibitors, vitronectin receptor antagonists, Src SH₂antagonists, vacular —H⁺-ATPase inhibitors, ipriflavone, fluoride,Tibolone, prostanoids, 17-beta hydroxysteroid dehydrogenase inhibitorsand Src kinase inhibitors.

Compounds of the present invention may be used in combination with malecontraceptives, such as nonoxynol 9 or therapeutic agents for thetreatment of hair loss, such as minoxidil and finasteride orchemotherapeutic agents, such as with LHRH agonists.

Further, the compounds of the present invention may be used incombination with anti-cancer and cytotoxic agents, including but notlimited to alkylating agents such as nitrogen mustards, alkylsulfonates, nitrosoureas, ethylenimines, and triazenes; antimetabolitessuch as folate antagonists, purine analogues, and pyrimidine analogues;antibiotics such as anthracyclines, bleomycins, mitomycin, dactinomycin,and plicamycin; enzymes such as L-asparaginase; farnesyl-proteintransferase inhibitors; 5α-reductase inhibitors; inhibitors of17β-hydroxysteroid dehydrogenase type 3; hormonal agents such asglucocorticoids, estrogens/antiestrogens, androgens/antiandrogens,progestins, and luteinizing hormone-releasing hormone antagonists,octreotide acetate; microtubule-disruptor agents, such as ecteinascidinsor their analogs and derivatives; microtubule-stabilizing agents such astaxanes, for example, paclitaxel (Taxol®), docetaxel (Taxotere®), andtheir analogs, and epothilones, such as epothilones A-F and theiranalogs; plant-derived products, such as vinca alkaloids,epipodophyllotoxins, taxanes; and topiosomerase inhibitors;prenyl-protein transferase inhibitors; and miscellaneous agents such ashydroxyurea, procarbazine, mitotane, hexamethylmelamine, platinumcoordination complexes such as cisplatin and carboplatin; and otheragents used as anti-cancer and cytotoxic agents such as biologicalresponse modifiers, growth factors; immune modulators and monoclonalantibodies. The compounds of the invention may also be used inconjunction with radiation therapy.

Representative examples of these classes of anti-cancer and cytotoxicagents include but are not limited to mechlorethamine hydrochloride,cyclophosphamide, chlorambucil, melphalan, ifosfamide, busulfan,carmustin, lomustine, semustine, streptozocin, thiotepa, dacarbazine,methotrexate, thioguanine, mercaptopurine, fludarabine, pentastatin,cladribin, cytarabine, fluorouracil, doxorubicin hydrochloride,daunorubicin, idarubicin, bleomycin sulfate, mitomycin C, actinomycin D,safracins, saframycins, quinocarcins, discodermolides, vincristine,vinblastine, vinorelbine tartrate, etoposide, etoposide phosphate,teniposide, paclitaxel, tamoxifen, estramustine, estramustine phosphatesodium, flutamide, buserelin, leuprolide, pteridines, diyneses,levamisole, aflacon, interferon, interleukins, aldesleukin, filgrastim,sargramostim, rituximab, BCG, tretinoin, irinotecan hydrochloride,betamethosone, gemcitabine hydrochloride, altretamine, and topoteca andany analogs or derivatives thereof.

Preferred member of these classes include, but are not limited to,paclitaxel, cisplatin, carboplatin, doxorubicin, carminomycin,daunorubicin, aminopterin, methotrexate, methopterin, mitomycin C,ecteinascidin 743, or porfiromycin, 5-fluorouracil, 6-mercaptopurine,gemcitabine, cytosine arabinoside, podophyllotoxin or podophyllotoxinderivatives such as etoposide, etoposide phosphate or teniposide,melphalan, vinblastine, vincristine, leurosidine, vindesine andleurosine.

Examples of anticancer and other cytotoxic agents include the following:epothilone derivatives as found in German Patent No. 4138042.8; WO97/19086, WO 98/22461, WO 98/25929, WO 98/38192, WO 99/01124, WO99/02224, WO 99/02514, WO 99/03848, WO 99/07692, WO 99/27890, WO99/28324, WO 99/43653, WO 99/54330, WO 99/54318, WO 99/54319, WO99/65913, WO 99/67252, WO 99/67253 and WO 00/00485; cyclin dependentkinase inhibitors as found in WO 99/24416 (see also U.S. Pat. No.6,040,321); and prenyl-protein transferase inhibitors as found in WO97/30992 and WO 98/54966; and agents such as those described genericallyand specifically in U.S. Pat. No. 6,011,029 (the compounds of which U.S.Patent can be employed together with any NHR modulators (including, butnot limited to, those of present invention) such as AR modulators, ERmodulators, with LHRH modulators, or with surgical castration,especially in the treatment of cancer).

The above other therapeutic agents, when employed in combination withthe compounds of the present invention, may be used, for example, inthose amounts indicated in the Physicians' Desk Reference (PDR) or asotherwise determined by one of ordinary skill in the art.

The compounds of the formula I can be administered for any of the usesdescribed herein by any suitable means, for example, orally, such as inthe form of tablets, capsules, granules or powders; sublingually;bucally; parenterally, such as by subcutaneous, intravenous,intramuscular, or intrastemal injection or infusion techniques (e.g., assterile injectable aqueous or non-aqueous solutions or suspensions);nasally, including administration to the nasal membranes, such as byinhalation spray; topically, such as in the form of a cream or ointment;or rectally such as in the form of suppositories; in dosage unitformulations containing non-toxic, pharmaceutically acceptable vehiclesor diluents. The present compounds can, for example, be administered ina form suitable for immediate release or extended release. Immediaterelease or extended release can be achieved by the use of suitablepharmaceutical compositions comprising the present compounds, or,particularly in the case of extended release, by the use of devices suchas subcutaneous implants or osmotic pumps. The present compounds canalso be administered liposomally.

Exemplary compositions for oral administration include suspensions whichcan contain, for example, microcrystalline cellulose for imparting bulk,alginic acid or sodium alginate as a suspending agent, methylcelluloseas a viscosity enhancer, and sweeteners or flavoring agents such asthose known in the art; and immediate release tablets which can contain,for example, microcrystalline cellulose, dicalcium phosphate, starch,magnesium stearate and/or lactose and/or other excipients, binders,extenders, disintegrants, diluents and lubricants such as those known inthe art. The compounds of formula I can also be delivered through theoral cavity by sublingual and/or buccal administration. Molded tablets,compressed tablets or freeze-dried tablets are exemplary forms which maybe used. Exemplary compositions include those formulating the presentcompound(s) with fast dissolving diluents such as mannitol, lactose,sucrose and/or cyclodextrins. Also included in such formulations may behigh molecular weight excipients such as celluloses (avicel) orpolyethylene glycols (PEG). Such formulations can also include anexcipient to aid mucosal adhesion such as hydroxy propyl cellulose(HPC), hydroxy propyl methyl cellulose (HPMC), sodium carboxy methylcellulose (SCMC), maleic anhydride copolymer (e.g., Gantrez), and agentsto control release such as polyacrylic copolymer (e.g. Carbopol 934).Lubricants, glidants, flavors, coloring agents and stabilizers may alsobe added for ease of fabrication and use.

Exemplary compositions for nasal aerosol or inhalation administrationinclude solutions in saline which can contain, for example, benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, and/or other solubilizing or dispersing agents such asthose known in the art.

Exemplary compositions for parenteral administration include injectablesolutions or suspensions which can contain, for example, suitablenon-toxic, parenterally acceptable diluents or solvents, such asmannitol, 1,3-butanediol, water, Ringer's solution, an isotonic sodiumchloride solution, or other suitable dispersing or wetting andsuspending agents, including synthetic mono- or diglycerides, and fattyacids, including oleic acid, or Cremaphor.

Exemplary compositions for rectal administration include suppositorieswhich can contain, for example, a suitable non-irritating excipient,such as cocoa butter, synthetic glyceride esters or polyethyleneglycols, which are solid at ordinary temperatures, but liquify and/ordissolve in the rectal cavity to release the drug.

Exemplary compositions for topical administration include a topicalcarrier such as Plastibase (mineral oil gelled with polyethylene).

The effective amount of a compound of the present invention can bedetermined by one of ordinary skill in the art, and includes exemplarydosage amounts for an adult human of from about 0.01 to 2000 mg ofactive compound per day, which can be administered in a single dose orin the form of individual divided doses, such as from 1 to 4 times perday. It will be understood that the specific dose level and frequency ofdosage for any particular subject can be varied and will depend upon avariety of factors including the activity of the specific compoundemployed, the metabolic stability and length of action of that compound,the species, age, body weight, general health, sex and diet of thesubject, the mode and time of administration, rate of excretion, drugcombination, and severity of the particular condition. Preferredsubjects for treatment include animals, most preferably mammalianspecies such as humans, and domestic animals such as dogs, cats and thelike, subject to NHR-associated conditions.

Transactivation Assays:

AR Specific Assay:

Compounds of the present invention were tested in transactivation assaysof a transfected reporter construct and using the endogenous androgenreceptor of the host cells. The transactivation assay provides a methodfor identifying functional agonists and partial agonists that mimic, orantagonists that inhibit, the effect of native hormones, in this case,dihydrotestosterone (DHT). This assay can be used to predict in vivoactivity as there is a good correlation in both series of data. See,e.g. T. Berger et al., J. Steroid Biochem. Molec. Biol. 773 (1992), thedisclosure of which is herein incorporated by reference.

For the transactivation assay a reporter plasmid is introduced bytransfection (a procedure to induce cells to take foreign genes) intothe respective cells. This reporter plasmid, comprising the cDNA for areporter protein, such as secreted alkaline phosphatase (SEAP),controlled by prostate specific antigen (PSA) upstream sequencescontaining androgen response elements (AREs). This reporter plasmidfunctions as a reporter for the transcription-modulating activity of theAR. Thus, the reporter acts as a surrogate for the products (mRNA thenprotein) normally expressed by a gene under control of the AR and itsnative hormone. In order to detect antagonists, the transactivationassay is carried out in the presence of constant concentration of thenatural AR hormone (DHT) known to induce a defined reporter signal.Increasing concentrations of a suspected antagonist will decrease thereporter signal (e.g., SEAP production). On the other hand, exposing thetransfected cells to increasing concentrations of a suspected agonistwill increase the production of the reporter signal.

For this assay, LNCaP and MDA 453 cells were obtained from the AmericanType Culture Collection (Rockville, Md.), and maintained in RPMI 1640 orDMEM medium supplemented with 10% fetal bovine serum (FBS; Gibco)respectively. The respective cells were transiently transfected byelectroporation according to the optimized procedure described byHeiser, 130 Methods Mol. Biol., 117 (2000), with thepSEAP2/PSA540/Enhancer reporter plasmid. The reporter plasmid, wasconstructed as follows: commercial human placental genomic DNA was usedto generate by Polymerase Cycle Reaction (PCR) a fragment containing theBglII site (position 5284) and the Hind III site at position 5831 of thehuman prostate specific antigen promoter (Accession #U37672), Schuur, etal., J. Biol. Chem., 271 (12): 7043-51 (1996). This fragment wassubcloned into the pSEAP2/basic (Clontech) previously digested withBglII and HindIII to generate the pSEAP2/PSA540 construct. Then afragment bearing the fragment of human PSA upstream sequence betweenpositions −5322 and −3873 was amplified by PCR from human placentalgenomic DNA. A XhoI and a BglII sites were introduced with the primers.The resulting fragment was subcloned into pSEAP2/PSA540 digested withXhoI and BglII respectively, to generate the pSEAP2/PSA540/Enhancerconstruct. LNCaP and MDA MB-453 cells were collected in media containing10% charcoal stripped FBS. Each cell suspension was distributed into twoGene Pulser Cuvettes (Bio-Rad) which then received 8 μg of the reporterconstruct, and electroporated using a Bio-Rad Gene Pulser at 210 voltsand 960 μ Faraday. Following the transfections the cells were washed andincubated with media containing charcoal stripped fetal bovine serum inthe absence (blank) or presence (control) of 1 nM dihydrotestosterone(DHT; Sigma Chemical) and in the presence or absence of the standardanti-androgen bicalutamide or compounds of the present invention inconcentrations ranging from 10⁻¹⁰ to 10⁻⁵ M (sample). Duplicates wereused for each sample. The compound dilutions were performed on a Biomek2000 laboratory workstation.

After 48 h, a fraction of the supernatant was assayed for SEAP activityusing the Phospha-Light Chemiluminescent Reporter Gene Assay System(Tropix, Inc). Viability of the remaining cells was determined using theCellTiter 96 Aqueous Non-Radioactive Cell Proliferation Assay (MTSAssay, Promega). Briefly, a mix of a tetrazolium compound(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt; MTS) and an electron coupling reagent (phenazinemethosulfate; PMS) are added to the cells. MTS (Owen's reagent) isbioreduced by cells into a formazan that is soluble in tissue culturemedium, and therefore its absorbance at 490 nm can be measured directlyfrom 96 well assay plates without additional processing. The quantity offormazan product as measured by the amount of 490 nm absorbance isdirectly proportional to the number of living cells in culture. For eachreplicate the SEAP reading was normalized by the Abs490 value derivedfrom the MTS assay. For the antagonist mode, the % Inhibition wascalculated as:% Inhibition=100×(1−[average control−average blank/averagesample−average blank])Data was plotted and the concentration of compound that inhibited 50% ofthe normalized SEAP was quantified (IC₅₀).For the agonist mode % Control was referred as the effect of the testedcompound compared to the maximal effect observed with the naturalhormone, in this case DHT, and was calculated as:% Control=100×average sample−average blank/average control−average blankData was plotted and the concentration of compound that activates tolevels 50% of the normalized SEAP for the control was quantified (EC₅₀).GR Specificity Assay:

The reporter plasmid utilized was comprised of the cDNA for the reporterSEAP protein, as described for the AR specific transactivation assay.Expression of the reporter SEAP protein was controlled by the mousemammary tumor virus long terminal repeat (MMTV LTR) sequences thatcontains three hormone response elements (HREs) that can be regulated byboth GR and PR see, e.g. G. Chalepakis et al., Cell, 53(3), 371 (1988).This plasmid was transfected into A549 cells, which expresses endogenousGR, to obtain a GR specific transactivation assay. A549 cells wereobtained from the American Type Culture Collection (Rockville, Md.), andmaintained in RPMI 1640 supplemented with 10% fetal bovine serum (FBS;Gibco). Determination of the GR specific antagonist activity of thecompounds of the present invention was identical to that described forthe AR specific transactivation assay, except that the DHT was replacedwith 5 nM dexamethasone (Sigma Chemicals), a specific agonist for GR.Determination of the GR specific agonist activity of the compounds ofthe present invention was performed as described for the ARtransactivation assay, wherein one measures the activation of the GRspecific reporter system by the addition of a test compound, in theabsence of a known GR specific agonists ligand.

PR Specific Assay:

The reporter plasmid utilized was comprised of the cDNA for the reporterSEAP protein, as described for the AR specific transactivation assay.Expression of the reporter SEAP protein was controlled by the mousemammary tumor virus long terminal repeat (MMTV LTR) sequences thatcontains three hormone response elements (HRE's) that can be regulatedby both GR and PR. This plasmid was transfected into T47D, whichexpresses endogenous PR, to obtain a PR specific transactivation assay.T47D cells were obtained from the American Type Culture Collection(Rockville, Md.), and maintained in DMEM medium supplemented with 10%fetal bovine serum (FBS; Gibco). Determination of the PR specificantagonist activity of the compounds of the present invention wasidentical to that described for the AR specific transactivation assay,except that the DHT was replaced with 1 nM Promegastone (NEN), aspecific agonist for PR. Determination of the PR specific agonistactivity of the compounds of the present invention was performed asdescribed for the AR transactivation assay, wherein one measures theactivation of the PR specific reporter system by the addition of a testcompound, in the absence of a known PR specific agonists ligand.

AR Binding Assay:

For the whole-cell binding assay, human LNCaP cells (T877A mutant AR) orMDA 453 (wild type AR) in 96-well microtiter plates containing RPMI 1640or DMEM supplemented with 10% charcoal stripped CA-FBS (CocalecoBiologicals) respectively, were incubated at 37° C. to remove anyendogenous ligand that might be complexed with the receptor in thecells. After 48 h, either a saturation analysis to determine the K_(d)for tritiated dihydrotestosterone, [³H]-DHT, or a competitive bindingassay to evaluate the ability of test compounds to compete with [³H]-DHTwere performed. For the saturation analysis, media (RPMI 1640 orDMEM—0.2% CA-FBS) containing [³H]-DHT (in concentrations ranging from0.1 nM to 16 nM) in the absence (total binding) or presence(non-specific binding) of a 500-fold molar excess of unlabeled DHT wereadded to the cells. After 4 h at 37° C., an aliquot of the total bindingmedia at each concentration of [³H]-DHT was removed to estimate theamount of free [³H]-DHT. The remaining media was removed, cells werewashed three times with PBS and harvested onto UniFilter GF/B plates(Packard), Microscint (Packard) was added and plates counted in aTop-Counter (Packard) to evaluate the amount of bound [³H]-DHT.

For the saturation analysis, the difference between the total bindingand the non-specific binding, was defined as specific binding. Thespecific binding was evaluated by Scatchard analysis to determine theK_(d) for [³H]-DHT. See e.g. D. Rodbard, Mathematics and statistics ofligand assays: an illustrated guide: In: J. Langon and J. J. Clapp,eds., Ligand Assay, Masson Publishing U.S.A., Inc., New York, pp. 45-99,(1981), the disclosure of which is herein incorporated by reference.

For the competition studies, media containing 1 nM [³H]-DHT andcompounds of the invention (“test compounds”) in concentrations rangingfrom 10⁻¹⁰ to 10⁻⁵ M were added to the cells. Two replicates were usedfor each sample. After 4 h at 37° C., cells were washed, harvested andcounted as described above. The data was plotted as the amount of[³H]-DHT (% of control in the absence of test compound) remaining overthe range of the dose response curve for a given compound. Theconcentration of test compound that inhibited 50% of the amount of[³H]-DHT bound in the absence of competing ligand was quantified (IC₅₀)after log-logit transformation. The K_(I) values were determined byapplication of the Cheng-Prusoff equation to the IC₅₀ values, where:

$K_{1} = {\frac{{IC}_{50}}{\left( {1 + {{\left( {}^{3}{H - {DHT}} \right)/K_{d}}\mspace{14mu}{{for}\mspace{14mu}}^{3}H} - {DHT}} \right)}.}$After correcting for non-specific binding, IC₅₀ values were determined.The IC₅₀ is defined as the concentration of competing ligand needed toreduce specific binding by 50%. The K_(d)s for [³H]-DHT for MDA 453 andLNCaP were 0.7 and 0.2 nM respectively.C2C12 Mouse Myoblast Transactivation Assay:

Two functional transactivation assays were developed to assess theefficacy of androgen agonists in a muscle cell background using aluciferase reporter. The first assay (ARTA Stable 1) uses a cell line,Stable 1 (clone #72), which expresses the full length rat androgenreceptor but requires the transient transfection of anenhancer/reporter. This cell line was derived from C2C12 mouse moyoblastcells. The second assay (ARTA Stable 2) uses a cell line, Stable 2(clone #133), derived from Stable 1 which expresses both rAR and theenhancer/luciferase reporter.

The enhancer/reporter construct used in this system ispGL3/2XDR-1/luciferase. 2XDR-1 was reported to be an AR specificresponse element in CV-1 cells, Brown et. al. The Journal of BiologicalChemistry 272, 8227-8235, (1997). It was developed by random mutagenesisof an AR/GR consensus enhancer sequence.

ARTA Stable 1:

-   1. Stable 1 cells are plated in 96 well format at 6,000 cells/well    in high glucose DMEM without phenol red (Gibco BRL, Cat. No.:    21063-029) containing 10% charcoal and dextran treated FBS (HyClone    Cat. No.: SH30068.02), 50 mM HEPES Buffer (Gibco BRL, Cat. No.:    15630-080), 1×MEM Na Pyruvate (Gibco BRL, Cat. No.: 11360-070), 0.5×    Antibiotic-Antimycotic, and 800 μg/mL Geneticin (Gibco BRL, Cat.    No.: 10131-035).-   2. 48 h later, cells are transfected with pGL3/2XDR-1/luciferase    using LipofectAMINE Plus™ Reagent (Gibco BRL, Cat. No.: 10964-013).    Specifically, 5 ng/well pGL3/2XDR-1/luciferase DNA and 50 ng/well    Salmon Sperm DNA (as carrier) are diluted with 5 □l/well Opti-MEMem    media (Gibco BRL, Cat. No.: 31985-070). To this, 0.5 □l/well Plus    reagent is added. This mixture is incubated for 15 min at rt. In a    separate vessel, 0.385 μl/well LipofectAMINE reagent is diluted with    5 □l/well Opti-MEM. The DNA mixture is then combined with the    LipofectAMINE mixture and incubated for an additional 15 min at rt.    During this time, the media from the cells is removed and replaced    with 60 μl/well of Opti-MEM. To this is added 10 □l/well of the    DNA/LipofectAMINE transfection mixture. The cells are incubated for    4 h.-   3. The transfection mixture is removed from the cells and replaced    with 90 μl of media as in #1 above.-   4. 10 μl/well of appropriate drug dilution is placed in each well.-   5. 24 h later, the Steady-Glo™ Luciferase Assay System is used to    detect activity according to the manufacturer's instructions    (Promega, Cat. No.: E2520).    ARTA Stable 2:-   1. Stable 2 cells are plated in 96 well format at 6,000 cells/well    in high glucose DMEM without phenol red (Gibco BRL, Cat. No.:    21063-029) containing 10% charcoal and dextran treated FBS (HyClone    Cat. No.: SH30068.02), 50 mM HEPES Buffer (Gibco BRL, Cat. No.:    15630-080), 1×MEM Na Pyruvate (Gibco BRL, Cat. No.: 11360-070), 0.5×    Antibiotic-Antimycotic, 800 μg/mL Geneticin (Gibco BRL, Cat. No.:    10131-035) and 800 μg/mL Hygromycin P (Gibco BRL, Cat. No.:    10687-010).-   2. 48 h later, the media on the cells is removed and replaced with    90 μl fresh. 10 μl/well of appropriate drug dilution is placed in    each well.-   3. 24 h later, the Steady-GloTM Luciferase Assay System is used to    detect activity according to the manufacturer's instructions    (Promega, Cat. No. E2520).    Proliferation Assays:    Human Prostate Cell Proliferation Assay:

Compounds of the present invention were tested (“test compounds”) on theproliferation of human prostate cancer cell lines. For that, MDA PCa2bcells, a cell line derived from the metastasis of a patient that failedcastration, Navone et al., Clin. Cancer Res., 3, 2493-500 (1997), wereincubated with or without the test compounds for 72 h and the amount of[³H]-thymidine incorporated into DNA was quantified as a way to assessnumber of cells and therefore proliferation. The MDA PCa2b cell line wasmaintained in BRFF-HPC1 media (Biological Research Faculty & FacilityInc., MD) supplemented with 10% FBS. For the assay, cells were plated inBiocoated 96-well microplates and incubated at 37° C. in 10% FBS(charcoal-stripped)/BRFF-BMZERO (without androgens). After 24 h, thecells were treated in the absence (blank) or presence of 1 nM DHT(control) or with test compounds (sample) of the present invention inconcentrations ranging from 10⁻¹⁰ to 10⁻⁵ M. Duplicates were used foreach sample. The compound dilutions were performed on a Biomek 2000laboratory work station. Seventy-two h later 0.44 uCi. of [³H]-Thymidine(Amersham) was added per well and incubated for another 24 h followed bytripsinization, harvesting of the cells onto GF/B filters. Micro-scintPS were added to the filters before counting them on a Beckman TopCount.

The % Inhibition was calculated as:%Inhibition=100×(1−[average_(control)−average_(blank)/average_(sample)−average_(blank)])Data was plotted and the concentration of compound that inhibited 50% ofthe [³H]-Thymidine incorporation was quantified (IC₅₀).Murine Breast Cell Proliferation Assay:

The ability of compounds of the present invention (“test compounds”) tomodulate the function of the AR was determined by testing said compoundsin a proliferation assay using the androgen responsive murine breastcell line derived from the Shionogi tumor, Hiraoka et al., Cancer Res.,47, 6560-6564 (1987). Stable AR dependent clones of the parentalShionogi line were established by passing tumor fragments under thegeneral procedures originally described in Tetuo, et. al., CancerResearch 25, 1168-1175 (1965). From the above procedure, one stableline, SC114, was isolated, characterized and utilized for the testing ofexample compounds. SC114 cells were incubated with or without the testcompounds for 72 h and the amount of [3H]-thymidine incorporated intoDNA was quantified as a surrogate endpoint to assess the number of cellsand therefore the proliferation rate as described in Suzuki et. al., J.Steroid Biochem. Mol. Biol. 37, 559-567 (1990). The SC114 cell line wasmaintained in MEM containing 10⁻⁸ M testosterone and 2% DCC-treated FCS.For the assay, cells were plated in 96-well microplates in themaintenance media and incubated at 37° C. On the following day, themedium was changed to serum free medium [Ham's F-12:MEM (1;1, v/v)containing 0.1% BSA] with (antagonist mode) or without (agonist mode)10⁻⁸ M testosterone and the test compounds of the present invention inconcentrations ranging from 10⁻¹⁰ to 10⁻⁵ M. Duplicates were used foreach sample. The compound dilutions were performed on a Biomek 2000laboratory work station. Seventy two h later 0.44 uCi of [3H]-Thymidine(Amersham) was added per well and incubated for another 2 h followed bytripsinization, and harvesting of the cells onto GF/B filters.Micro-scint PS were added to the filters before counting them on aBeckman TopCount.

For the antagonist mode, the % Inhibition was calculated as:%Inhibition=100×(1−[average_(sample)−average_(blank)/average_(control)−average_(blank)])Data was plotted and the concentration of compound that inhibited 50% ofthe [³H]-Thymidine incorporation was quantified (IC₅₀).For the agonist mode % Control was referred as the effect of the testedcompound compared to the maximal effect observed with the naturalhormone, in this case DHT, and was calculated as:%Control=100×(average_(sample)−average_(blank))/(average_(control)−average_(blank))Data was plotted and the concentration of compound that inhibited 50% ofthe [³H]-Thymidine incorporation was quantified (EC₅₀).In Vitro Assay to Measure GR-Induced AP-1 Transrepression:

The AP-1 assay is a cell-based luciferase reporter assay. A549 cells,which contain endogenous glucocorticoid receptor, were stablytransfected with an AP-1 DNA binding site attached to the luciferasegene. Cells are then grown in RPMI+10% fetal calf serum(charcoal-treated)+Penicillin/Streptomycin with 0.5 mg/mL geneticin.Cells are plated the day before the assay at approximately 40000cells/well. On assay day, the media is removed by aspiration and 20 μLassay buffer (RPMI without phenol red+10% FCS(charcoal-treated)+Pen/Strep) is added to each well. At this pointeither 20 μL assay buffer (control experiments), the compounds of thepresent invention (“test compounds”) (dissolved in DMSO and added atvarying concentrations) or dexamethasome (100 nM in DMSO, positivecontrol) are added to each well. The plates are then pre-incubated for15 min at 37° C., followed by stimulation of the cells with 10 ng/mLPMA. The plates are then incubated for 7 h at 37° C. after which 40 μLluciferase substrate reagent is added to each well. Activity is measuredby analysis in a luminometer as compared to control experiments treatedwith buffer or dexamethasome. Activity is designated as % inhibition ofthe reporter system as compared to the buffer control with 10 ng/mL PMAalone. The control, dexamethasone, at a concentration of <10 □Mtypically suppresses activity by 65%. Test compounds which demonstratean inhibition of PMA induction of 50% or greater at a concentration oftest compound of ≦10 □M are deemed active.

In Vivo Assays

Levator Ani & Wet Prostate Weight Assay AR Agonist Assay:

The activity of compounds of the present invention as AR agonists wasinvestigated in an immature male rat model, a recognized test ofanabolic effects in muscle and sustaining effects in sex organs for agiven compound, as described in L. G. Hershberger et al., Proc. Soc.Expt. Biol. Med., 83, 175 (1953); B. L. Beyler et al, “Methods forevaluating anabolic and catabolic agents in laboratory animals”, J.Amer. Med. Women's Ass., 23, 708 (1968); H. Fukuda et al.,“Investigations of the levator ani muscle as an anabolic steroid assay”,Nago Dai. Yak. Ken. Nem. 14, 84 (1966) the disclosures of which areherein incorporated by reference.

The basis of this assay lies in the well-defined action of androgenicagents on the maintenance and growth of muscle tissues and sexualaccessory organs in animals and man. Androgenic steroids, such astestosterone (T), have been well characterized for their ability tomaintain muscle mass. Treatment of animals or humans after castrationswith an exogenous source of T results in a reversal of muscular atrophy.The effects of T on muscular atrophy in the rat levator ani muscle havebeen well characterized. M. Masuoka et al., “Constant cell population innormal, testosterone deprived and testosterone stimulated levator animuscles” Am. J. Anat. 119, 263 (1966); Z. Gori et al., “Testosteronehypertrophy of levator ani muscle of castrated rats. I. Quantitativedata” Boll.—Soc. Ital. Biol. Sper. 42, 1596 (1966); Z. Gori et al.,“Testosterone hypertrophy of levator ani muscle of castrated rats. II.Electron-microscopic observations” Boll.—Soc. Ital. Biol. Sper. 42, 1600(1966); A. Boris et al., Steroids 15, 61 (1970). As described above, theeffects of androgens on maintenance of male sexual accessory organs,such as the prostate and seminal vesicles, is well described. Castrationresults in rapid involution and atrophy of the prostate and seminalvesicles. This effect can be reversed by exogenous addition ofandrogens. Since both the levator ani muscle and the male sex organs arethe tissues most responsive to the effects of androgenic agents, thismodel is used to determine the androgen dependent reversal of atrophy inthe levator ani muscle and the sex accessory organs in immaturecastrated rats. Sexually mature rats (200-250 g, 6-8 weeks-old,Sprague-Dawley, Harlan) were acquired castrated from the vendor(Taconic). The rats were divided into groups and treated daily for 7 to14 days with one of the following:

-   -   1. Control vehicle    -   2. Testosterone Propionate (TP) (3 mg/rat/day, subcutaneous)    -   3. TP plus Bicalutamide (administered p.o. in PEGTW, QD), a        recognized antiandrogen, as a reference compound.    -   4. To demonstrate antagonist activity, a compound of the present        invention (“test compound”) was administered (p.o. in PEGTW, QD)        with TP (s.c. as administered in group 2) in a range of doses.    -   5. To demonstrate agonist activity a compound of the present        invention (“test compound”) was administered alone (p.o. in        PEGTW, QD) in a range of doses.

At the end of the 7-14-day treatment, the animals were sacrificed bycarbon dioxide, and the levator ani, seminal vesicle and ventralprostate weighed. To compare data from different experiments, thelevator ani muscle and sexual organ weights were first standardized asmg per 100 g of body weight, and the increase in organ weight induced byTP was considered as the maximum increase (100%). Super-anova (onefactor) was used for statistical analysis.

The gain and loss of sexual organ weight reflect the changes of the cellnumber (DNA content) and cell mass (protein content), depending upon theserum androgen concentration. See Y. Okuda et al., J. Urol., 145,188-191 (1991), the disclosure of which is herein incorporated byreference. Therefore, measurement of organ wet weight is sufficient toindicate the bioactivity of androgens and androgen antagonist. Inimmature castrated rats, replacement of exogenous androgens increaseslevator ani, seminal vesicles (SV) and prostate in a dose dependentmanner.

The maximum increase in organ weight was 4 to 5-fold when dosing 3mg/rat/day of testosterone (T) or 1 mg/rat/day of testosteronepropionate (TP) for 3 days. The EC₅₀ of T and TP were about 1 mg and0.03 mg, respectively. The increase in the weight of the VP and SV alsocorrelated with the increase in the serum T and DHT concentration.Although administration of T showed 5-times higher serum concentrationsof T and DHT at 2 h after subcutaneous injection than that of TP,thereafter, these high levels declined very rapidly. In contrast, theserum concentrations of T and DHT in TP-treated animals were fairlyconsistent during the 24 h, and therefore, TP showed about 10-30-foldhigher potency than free T.

The following examples serve to better illustrate, but not limit, someof the preferred embodiments of the invention.

EXAMPLE 1 (2S,3R)-3-hydroxy-2-pyrrolidinecarboxylic acid methyl ester

1A. (2S,3S)-3-Hydroxy-2-pyrrolidinecarboxylic acid methyl esterhydrochloride salt

Hydrogen chloride gas was bubbled through a suspension oftrans-3-hydroxy-L-proline (50 g, 0.38 mol) in MeOH (600 mL) cooled to 0°C. for 10 min. The resulting clear solution was stirred at rt for 4 h,then concentrated carefully under reduced pressure (white precipitatesformed during the concentration). The resulting white solid was driedunder vacuum overnight to afford the title compound (68.26 g) as a whitesolid.

1B. (2S,3S)-N-tert-Butyloxycarbonyl-3-hydroxy-2-pyrrolidinecarboxylicacid methyl ester

To a suspension of compound 1A (68.26 g, 0.375 mol) in CH₂Cl₂ (1.0 L)cooled to 0° C. was added Et₃N (105.3 mL, 0.755 mol), followed byportionwise addition of di-tert-butyl dicarbonate (82.96 g, 0.380 mol).The resulting mixture was stirred at rt for 4 h, then partitionedbetween water and CH₂Cl₂. The CH₂Cl₂ layer was washed with water (2×),20% aqueous citric acid (1×), water, brine, dried (Na₂SO₄) andconcentrated under reduced pressure to give an oily residue. The crudeproduct was chromatographed (silica gel) eluting with 15%-50%EtOAc/hexane to afford compound 1B (73.3 g) as a pale yellow viscousoil.

1C.(2S,3R)-N-tert-Butyloxycarbonyl-3-benzoyloxy-2-pyrrolidine-carboxylicacid methyl ester

To a stirred solution of compound 1B (69.1 g, 0.282 mol), Ph₃P (88.7 g,0.338 mol) and benzoic acid (41.32 g, 0.338 mol) in anhydrous THF (1.35L) cooled to 0° C. was added a solution of DEAD (62 mL, 0.33 mol) inanhydrous THF (50 mL) dropwise over 1 h through an addition funnel.After the addition, the resulting light yellow solution was stirred atrt until the reaction was complete (˜8 h). The reaction mixture was thenpartitioned between EtOAc and aqueous NaHCO₃. The organic layer waswashed with saturated aqueous NaHCO₃, water (2×), brine, dried (Na₂SO₄)and concentrated under reduced pressure to yield a crude product as asemi-solid. The crude product was suspended in 25% EtOAc/hexane andstirred vigorously for 3 h. The resulting suspension was filtered andthe collected white solid (triphenylphosphine oxide) rinsed with 20%EtOAc/hexane (2×). The combined filtrate was concentrated under reducedpressure to yield an oily residue, which was triturated twice with 20%EtOAc/hexane as described above to yield approximately 150 g of thepartially purified product as a yellow oil, which was further purifiedby flash chromatography (silica gel) eluting with 10-20% EtOAc/hexane tofurnish pure compound 1C (88.4 g) as a light yellow viscous oil.

1D. (2S,3R)-N-tert-Butyloxycarbonyl-3-hydroxy-2-pyrrolidinecarboxylicacid methyl ester

To a solution of compound 1C (88.44 g, 0.253 mol) in anhydrous MeOH (700mL) cooled to 0° C. was slowly added a freshly prepared 1N solution ofKOH in anhydrous MeOH (367 mL, 0.367 mol) over 25 min through anaddition funnel. After the addition, the light yellow solution wasstirred at 0° C. for 2 h, and then the reaction was quenched by slowaddition (over 25 min) of a solution of 1N HCl in dioxane/EtOAc (380 mL)through an addition funnel. The resulting white suspension wasconcentrated under reduced pressure to remove most of the solvent, andthe remaining mixture was partitioned between water and EtOAc. Theseparated organic phase was washed with water (2×), saturated aqueousNaHCO₃ (2×), water, brine, and then dried (Na₂SO₄). The filtrate wasconcentrated under reduced pressure to give a light yellow oily residue,which was chromatographed (silica gel) eluting first with 25-30%EtOAc/hexane, then 5% MeOH in 30% EtOAc/hexane to furnish compound 1D(44.6 g) as a pale yellow oil.

1E. (2S,3R)-3-Hydroxy-2-pyrrolidinecarboxylic acid methyl ester,trifluoroacetic acid salt

To a solution of compound 1D (44.6 g, 0.182 mol) in CH₂Cl₂ (450 mL)cooled to 0° C. was slowly added TFA (275 mL) through an addition funnelover 40 min. After addition, the reaction mixture was stirred at 0° C.for 2 h, then concentrated under reduced pressure to give a viscous oilyresidue which was evaporated with ether (2×), toluene (1×), ether (2×)and dried under vacuuum overnight to yield compound 1E (59.5 g) as alight yellow solid.

1F. (2S,3R)-3-Hydroxy-2-pyrrolidinecarboxylic acid methyl ester

To a solution of compound 1E (12.64 g, 48.8 mmol) in MeOH (150 mL) wasadded WA21J resin (60 g). The resulting suspension was stirred at rt for1 h, and then filtered. The collected resin was rinsed with MeOH (2×)and combined filtrate concentrated carefully under reduced pressure togive compound 1F (7.7 g) as a colorless oil. [□]_(D)=14.9° (c. 1.0,MeOH); HPLC: 100% at 0.157 min (retention time) (Conditions: Phenom.Luna C18 (4.6×50 mm); Eluted with 0% to 100% B; 4 min gradient (A=90%H₂O—10% MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄), Flow rate at4 mL/min., UV detection at 220 nm); MS (ES) m/z 146 [M+1]⁺

EXAMPLE 2(7R,7aS)-4-(7-Hydroxy-1,3-dioxo-tetrahydropyrrolo[1,2-c]imidazol-2-yl)-5,6,7,8-tetrahydronaphthalene-1-carbonitrile

2A. N-(5,6,7,8-Tetrahydronaphthalen-1-yl)-acetamide

To a solution of commercially available 5,6,7,8-tetrahydronaphthylamine(2 g, 14 mmol) in EtOH (5 mL) at rt was slowly added acetic anhydride(1.28 mL, 13.6 mmol). After addition, the reaction mixture was stirredat rt for 5 min. The resulting suspension was filtered, the collectedsolid washed with hexane (5×) and dried under vacuum to furnish thetitle compound (2.5 g) as an off-white solid.

2B. N-(4-Bromo-5,6,7,8-tetrahydronaphthalen-1-yl)-acetamide

To a solution of compound 2A (2 g, 11 mmol) in AcOH (15 mL) cooled at 0°C. was added a solution of bromine (1.69 mL, 33 mmol) in AcOH (1.2 mL)slowly so that the reaction temperature was maintained below 17° C.After addition, the reaction mixture was stirred at rt until all thestarting material was consumed (˜4 h). The reaction mixture was thenpoured into ice/water and the resulting suspension filtered. Thecollected solid was washed with H₂O until the filtrate pH=6-7, and driedin a vacuum oven at 50° C. overnight to yield compound 2B (2.7 g) as anoff-white solid.

2C. N-(4-Cyano-5,6,7,8-tetrahydronaphthalen-1-yl)-acetamide

A suspension of compound 2B (1 g, 3.7 mmol) and CuCN (0.335 g, 0.374mmol) in anhydrous DMF (8 mL) was refluxed for 5 h. After cooling to rt,the reaction mixture was concentrated under reduced pressure to removemost of the DMF and the remaining residue triturated with EtOAc (5×).The combined EtOAc was washed with water and brine, dried (Na₂SO₄), andconcentrated under reduced pressure to dryness to yield compound 2C (0.8g) as a yellow solid.

2D. 4-Amino-5,6,7,8-tetrahydronaphthalene-1-carbonitrile, hydrochloridesalt

A suspension of 2C (0.40 g, 1.87 mmol) in a mixed solvent of EtOH (2 mL)and conc. HCl (2 mL) was refluxed for 2 h. The resulting solution wasallowed to cool to rt and concentrated under reduced pressure. Theobtained solid was evaporated with toluene (2×) to yield compound 2D(0.25 g) as a solid.

2E. 4-Isocyanato-5,6,7,8-tetrahydronaphthalene-1-carbonitrile

To a stirring suspension of compound 2D (0.10 g, 0.48 mmol) and NaHCO₃(0.404 g, 4.80 mmol) in CH₂Cl₂ (4 mL) cooled to 0° C. was rapidly addeda solution of phosgene (20%) in toluene (0.95 mL, 4.32 mmol). Afteraddition, the mixture was stirred at rt for 2 h, then filtered to removethe solid. The filtrate was concentrated under reduced pressure, theresulting solid residue dried under vacuum for 1 h to afford compound 2E(95 mg) as a light yellow solid.

2F.(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-5,6,7,8-tetrahydronaphthalene-1-carbonitrile

To a suspension of compound 1E (0.145 g, 0.56 mmol) in CH₂Cl₂ (2 mL)cooled at 0° C. was added i-Pr₂NEt (0.12 mL, 0.69 mmol). After stirringat 0° C. for 20 min, compound 2E (95 mg, 0.48 mmol) in CH₂Cl₂ (1 mL)solution was added, along with 4 Å molecular sieves (0.5 g) and theresulting mixture stirred at rt until urea formation was complete (˜2h). To the mixture was then added DBU (0.15 mL, 1.0 mmol), the resultingbrown colored suspension was stirred vigorously at rt until hydantoinformation was complete (˜15 h). The reaction mixture was loaded on asilica gel column, eluted with 40% EtOAc/hexane, and 5% MeOH inEtOAc/hexane (1:1) to afford 128 mg of the title compound as a whitesolid. HPLC: 99% at 2.42 min (retention time) (Conditions: Phenom. LunaC18 (4.6×50 mm); Eluted with 0% to 100% B; 4 min gradient (A=90% H₂O—10%MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄ ), Flow rate at 4mL/min., UV detection at 220 nm). Chiral HPLC: retention time=9.01 min(99%); Conditions: (CHIRALPAK® OD column 4.6×250 mm; 25% isopropanol inhexane over 30 min at flow rate 1.0 mL/min, UV detection at 220 nm); MS(ES) m/z 312 [M+1]⁺

EXAMPLE 3(7R,7aS)-7-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-indan-4-carbonitrile

3A. N-Indan-4-yl-acetamide

A suspension of commercially available 4-nitroindane (10 g, 61.3 mmol)and 10% Pd/C (1 g) in MeOH (200 mL) was stirred vigorously under anatmosphere of hydrogen overnight at rt. The reaction was filteredthrough a pad of Celite® and concentrated, and residual solvent wasremoved by combining the reaction with toluene followed by evaporation.The crude reaction was taken up in pyridine (100 mL), and Ac₂O (20 mL)was added and the reaction stirred for 16 h at rt. The reaction wasevaporated and the crude product was purified by flash chromatography(5% MeOH in EtOAc/hexane(1:1) to give the title compound (9.12 g).

3B.(7R,7aS)-7-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)indan-4-carbonitrile

Compound 3B as a white solid was prepared from compound 3A by proceduresanalogous to those described in Example 2. HPLC: 97% at 2.75 min(retention time) (Conditions: YMC S5 ODS (4.6×50 mm); Eluted with 0% to100% B; 4 min gradient; (A=90% H₂O—10% MeOH—0.1% H₃PO₄ and B=10% H₂O—90%MeOH—0.1% H₃PO₄); Flow rate at 4 mL/min. UV detection at 220 nm). LC/MSm/z 298 [M+1]⁺.

EXAMPLE 4(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2,3-dihydrobenzofuran-7-carbonitrile

4A. 4-Acetylamino-2,3-dihydrobenzofuran-7-carboxylic acid amide

Ammonia gas was bubbled through a solution of4-acetylamino-2,3-dihydro-benzofuran-7-carboxylic acid methyl ester (12g, 51 mmol, prepared as described in Takuji Kakigami et al, Chem. Pharm.Bull, 46 (1), 42-52, (1998)) in CH₃OH (200 mL) in a pressure bottle at0-5° C. until the solution was saturated. The bottle was sealed andstirred at 60° C. overnight. After cooling to 0-5° C., the reactionmixture was charged one more time with NH₃ gas, then the sealed bottlestirred at 60° C. for another 24 h. After cooling to rt, whiteprecipitates were collected by filtration, and dried under vacuum tofurnish the title compound (10.5 g) as a white solid.

4B. N-(7-Cyano-2,3-dihydrobenzofuran-4-yl)-acetamide

To a suspension of compound 4A (3.5 g, 16 mmol) in THF (64 mL) at 0° C.under Ar was added pyridine (6.5 mL, 80 mmol), followed bytrifluoroacetic anhydride (5.6 mL, 40 mmol) dropwise. After addition,the reaction mixture was stirred at 0° C. for 5 min, then pored into H₂O(50 mL), extracted with EtOAc (3×100 mL). The combined EtOAc extractswere washed with brine, dried (Na₂SO₄), and concentrated under reducedpressure to give a crude product, which was purified by crystallizationfrom MeOH-EtOAc-Hexane to furnish the title compound (2.3 g) as a whitesolid.

4C.(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2,3-dihydrobenzofuran-7-carbonitrile

The title compound as a white solid was prepared from compound 4B byprocedures analogous to those described in Example 2 (from 2D to 2F).HPLC: 100% at 3.43 min (retention time) (Conditions: Zorbax SB C18(4.6×75 mm); Eluted with 0% to 100% B, 8 min gradient (A=90% H₂O—10%MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄). Flow rate at 2.5mL/min. UV detection at 220 nm.). Chiral HPLC: retention time=13.69 min(100%); Conditions: OD (4.6×250 mm); Eluted with 25% isopropanol inhexane for 30 min at 1 mL/min. MS (ES) m/z 300 [M+1]⁺.

EXAMPLE 5(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-chroman-8-carbonitrile

5A. 4-Acetylamino-2-hydroxy-3-(3-hydroxypropyl)-benzoic acid, methylester

To a suspension of 4-acetylamino-3-allyl-2-hydroxy-benzoic acid methylester (2.49 g, 10 mmol, prepared as described in Takuji Kakigami et.al., Chem. Pharm. Bull, 46 (1), 42-52, (1998)) in THF (10 mL) at 0° C.under Argon was added a 0.5 M solution of 9-BBN in THF (80 mL, 40 mmol).The reaction mixture was stirred at 0° C. for 40 min, then at rt for 2h. The reaction mixture was then cooled to 0° C., and 1 N aqueous NaOH(25 mL) was added dropwise over 5 min, followed by 30% aqueous H₂O₂ (20mL) over 5 min. After addition, the reaction mixture was stirred at rtfor 2 h, then extracted with EtOAc (3×60 mL). The combined EtOAcextracts were washed with brine, dried (Na₂SO₄), and concentrated underreduced pressure to give a crude product, which was chromatographed(silica gel) eluting with 30% to 80% EtOAc/hexane to afford the titlecompound (1.82 g) as a foam.

5B. 5-Acetylaminochroman-8-carboxylic acid, methyl ester

To a solution of compound 5A (1.80 g, 6.7 mmol) and Ph₃P (1.94 g, 7.4mmol) in THF (30 mL) cooled at 0° C. was added dropwise DEAD (1.17 mL,7.4 mmol). After addition, the reaction mixture was stirred at rt for 2h, then concentrated under reduced pressure. The residue waschromatographed (silica gel) eluting with 50% to 100% EtOAc/hexane tofurnish the title compound (1.3 g) as a white solid.

5C.(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)chroman-8-carbonitrile

The title compound was prepared from compound 5B and isolated as a whitesolid by procedures analogous to those described in Example 4. HPLC:100% at 3.58 min (retention time) (Conditions: Zorbax SB C18 (4.6×75mm); Eluted with 0% to 100% B, 8 min gradient. (A=90% H₂O—10% MeOH—0.1%H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄). Flow rate at 2.5 mL/min. UVdetection at 220 nm.). Chiral HPLC: retention time=12.35 min (99%);Conditions: OD (4.6×250 mm); Eluted with 25% isopropanol in hexane for30 min at 1 mL/min. MS (ES) m/z 314 [M+1]⁺.

EXAMPLE 6(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2,2-dimethyl-2H-chromene-8-carbonitrile

6A. 2-(1,1-Dimethylprop-2-ynyloxy)-4-nitrobenzonitrile

To a solution of commercially available 2-methyl-3-butyn-2-ol (0.28 mL,2.9 mmol) in anhydrous CH₃CN (1.5 mL) cooled to −5° C. was added DBU(0.56 mL, 3.7 mmol) followed by addition of trifluoroacetic anhydride(0.41 mL, 2.9 mmol) over 25 min while maintaining the reactiontemperature below 2° C. After the addition, the reaction mixture wasstirred at 0° C. for 30 min before using in the following reaction.

To a solution of 2-hydroxy-4-nitro-benzonitrile (413.6 mg, 2.52 mmol,prepared as described in Yasubiro Imakura et al, Chem. Pharm. Bull, 40(7), 1691-1696, (1992)) in CH₃CN (1.5 mL) cooled to −5° C. was added DBU(0.48 mL, 3.2 mmol) and CuCl₂.2H₂O (2 mg), followed by addition of asolution of trifluoroacetate prepared above over 30 min whilemaintaining the reaction temperature below 0° C. After addition, thereaction mixture was stirred at 0° C. for 2 h, then concentrated underreduced pressure. The residue was partitioned between EtOAc (100 mL) andwater (30 mL), and the separated organic layer washed with 1 N aqueousHCl (2×20 mL), 1 N aqueous NaOH (20 mL), 1 N aqueous NaHCO₃, brine, thendried (Na₂SO₄), and concentrated under reduced pressure. The crudeproduct was chromatographed (silica gel) eluting with 0% to 50%EtOAc/hexane to furnish the title compound (280 mg, 48%).

6B. 2,2-Dimethyl-5-nitro-2H-chromene-8-carbonitrile

A solution of compound 6A (270 mg, 1.17 mmol) in N,N-diethylaniline (1mL) was heated to 185° C. for 3 h. After cooling to rt, the reactionmixture was chromatographed (silica gel) eluting with 0-50% EtOAc/hexaneto afford the title compound (230 mg).

6C. 5-Amino-2,2-dimethyl-2H-chromene-8-carbonitrile

To a stirred solution of compound 6B (230 mg, 1 mmol) in EtOAc (5 mL)was added SnCl₂ 2H₂O (780 mg, 3.5 mmol). The resulting mixture wasstirred at rt for 20 h, then saturated aqueous K₂CO₃ was added. Afterstirring for 30 min, the reaction was treated with solid K₂CO₃ (550 mg).The resulting suspension was stirred at rt for another 2 h, and thenfiltered. The filtrate was concentrated under reduced pressure to give acrude product, which was chromatographed (silica gel) eluting with 20%to 60% EtOAc/hexane to afford the title compound (160 mg) as a lightyellow oil.

6D.(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2,2-dimethyl-2H-chromene-carbonitrile

The title compound was prepared from compound 6C by procedures analogousto those described in Example 2 (2E to 2F). HPLC: 100% at 4.48 min(retention time) (Conditions: Zorbax SB C18 (4.6×75 mm); Eluted with 0%to 100% B, 8 min gradient. (A=90% H₂O—10% MeOH—0.1% H₃PO₄ and B=10%H₂O—90% MeOH—0.1% H₃PO₄). Flow rate at 2.5 mL/min. UV detection at 220nm). Chiral HPLC: retention time=9.37 min (99%); Conditions: OD (4.6×250mm); Eluted with 25% isopropanol in hexane for 30 min at 1 mL/min. MS(ES) m/z 340 [M+1]⁺.

EXAMPLE 7(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2,2-dimethylchroman-8-carbonitrile

A suspension of compound 6D (50 mg, 0.15 mmol) and 5% Pd/C (10 mg) inEtOH (3 mL) was stirred at rt under an atmosphere of hydrogen for 2 h.The reaction mixture was then filtered, and the filtrate concentratedunder reduced pressure to give a crude product, which was purified bycrystallization from hot CH₂Cl₂-hexane to give the title compound (36mg) as a white solid. HPLC: 99% at 4.52 min (retention time)(Conditions: Zorbax SB C18 (4.6×75 mm); Eluted with 0% to 100% B, 8 mingradient. (A=90% H₂O—10% MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1%H₃PO₄). Flow rate at 2.5 mL/min. UV detection at 220 nm.). Chiral HPLC:retention time=8.72 min (99%); Conditions: OD (4.6×250 mm); Eluted with25% isopropanol in hexane for 30 min at 1 mL/min. MS (ES) m/z 342[M+1]⁺.

EXAMPLE 8(7R,7aS)-8-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2,3-dihydrobenzo[1,4]dioxane-5-carbonitrile

8A. 3-Nitrocatechol

To a solution of catechol (5 g, 45 mmol) in Et₂O (187 mL) cooled to 0°C. was added dropwise fuming HNO₃ (2 mL). After addition, the reactionwas allowed to stand at rt overnight, and the Et₂O was removed byevaporation under reduced pressure. The residue was triturated withpentane (3×), and the combined organics were dried (Na₂SO₄), filtered,and concentrated under reduced pressure. The residue was chromatographed(silica gel) eluting with 10%-20% EtOAc/hexane to give the titlecompound (2.94 g).

8B. 5-Nitrobenzo[1,4]dioxane

To a solution of compound 8A (3.0 g, 19.4 mmol) in DMF (40 mL) was addedCsF (14.7 g, 96.8 mmol), followed by dibromoethane (1.84 mL, 21.3 mmol).The mixture was heated to 110° C. for 1.5 h, then cooled to rt,partitioned between water and Et₂O. The separated Et₂O layer was washedwith water, saturated aqueous NaHCO₃, brine, dried (Na₂SO₄), filteredand concentrated under reduced pressure. The residue was chromatographed(silica gel) eluting with 5% MeOH in CH₂Cl₂ to afford the title compound(0.73 g).

8C. N-(Benzo[1,4]dioxan-5-yl)-acetamide

Compound 8C was prepared from 8B by procedures analogous to thosedescribed in Experiment 3A and 2A.

8D. N-(8-Bromobenzo[1,4]dioxan-5-yl)-acetamide

To a solution of compound 8C (0.80 g, 4.15 mmol) in chloroform (2.4 mL)cooled to −20° C. was slowly added a solution of bromine (0.22 mL, 4.35mmol) in chloroform (1 mL) so that the reaction temperature wasmaintained below −10° C. The reaction was stirred at 0° C. for 5 min,and then quenched immediately with water. The mixture was extracted withCH₂Cl₂ (3×), the combined extracts washed with saturated NaHCO₃, brine,dried (Na₂SO₄), filtered and concentrated under reduced pressure. Theresidue was chromatographed (silica gel) eluting with CH₂Cl₂ to affordthe title compound (0.98 g).

8E.(7R,7aS)-8-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzo[1,4]dioxane-5-carbonitrile

The title compound (32 mg) was prepared from compound 8D and isolated asa white solid by procedures analogous to those described in Example 2(2C to 2F). mp196-197° C.; HPLC: 99% at 12.31 min (retention time)(CHIRALPAK® OD column 4.6×250 mm; 25% isopropanol in hexane over 30 min,1 mL/min, UV detection at 220 nm); MS (ES) m/z 322 [M+1]⁺.

EXAMPLE 9(7R,7aS)-7-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methyl-3H-benzimidazole-4-carbonitrile

9A. 1-Methyl-1H-benzimidazol-4-ylamine

A suspension of 1-methyl-4-nitro-1H-benzimidazole [2.1 g, 12 mmol,prepared as described in Viktor Milata et. al., Org. Prep. Proced. Int.25 (6), 703-704 (1993)] and 5% Pd/C (0.21 g) in EtOH (40 mL) wasvigorously stirred under an atmoshphere of hydrogen at rt overnight. Thereaction mixture was filtered and the filtrate concentrated underreduced pressure to afford the tiltle compound (1.65 g).

9B. 7-Amino-3-methyl-3H-benzimidazole-4-carbonitrile

The title compound was prepared from compound 9A by procedures analogousto those described in Experiment 2 (2A to 2D).

9C. 7-Isocyanato-3-methyl-3H-benzimidazole-4-carbonitrile

To a suspension of compound 9B (240 mg, 1.39 mmol) in CH₂Cl₂ (10 mL)under Argon was added Et₃N (0.97 mL, 6.95 mmol). The resultingsuspension was stirred at 0° C. for 15 min, then a solution of phosgene(20%) in toluene (1.4 mL, 2.8 mmol) was added. After the addition, themixture was stirred at rt for 2 h, then concentrated under reducedpressure. The resulting solid residue was dried under vacuum for 1 h toafford the title compound, which was used immediately in the preparationof compound 9D.

9D.(3R,2aS)-1-(7-Cyano-1-methyl-1H-benzimidazol-4-ylcarbamoyl)-3-hydroxypyrrolidine-2-carboxylicacid methyl ester

To a suspension of 1E (600 mg, 1.67 mmol) in CH₂Cl₂ (4 mL) at 0° C. wasadded i-Pr₂NEt (0.35 mL, 2.0 mmol). After 5 min, a suspension ofcompound 9C (1.37 mmol) in CH₂Cl₂ (3 mL) was added, followed by 4 Åmolecular sieves (˜1.0 g). The resulting mixture was stirred at rt for 3h, and then concentrated under reduced pressure. The crude product waschromatographed (silica gel) eluting with 4-6% MeOH in CH₂Cl₂ to affordthe title compound (250 mg) as a light yellow solid.

9E.(7R,7aS)-7-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methyl-3H-benzoimidazole-4-carbonitrile

To a suspension of compound 9D (51.5 mg, 0.15 mmol) in1,2-dichloroethane (1 mL) at rt was added DBU (34 μL, 0.23 mmol), alongwith 4 Å molecular sieves (˜0.2 g). The resulting mixture was stirred atrt for 20 h, then concentrated under reduced pressure. The crude productwas chromatographed (silica gel) eluting with 4-6% MeOH in CH₂Cl₂ togive the title compound (25 mg) as a white solid. HPLC: 98% at 2.52 min(retention time) (Conditions: Zorbax SB C18 (4.6×75 mm); Eluted with 0%to 100% B, 8 min gradient. (A=90% H₂O—10% MeOH—0.1% H₃PO₄ and B=10%H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5 mL/min. UV detection at 220nm.). Chiral HPLC: retention time=62.5 min (98%); Conditions: OD(4.6×250 mm); Eluted with 45% isopropanol in hexane for 90 min at 1mL/min. MS (ES) m/z 312 [M+1]⁺.

EXAMPLE 104-(7-Hydroxy-1,3-dioxotetrahydro-pyrrolo[1,2-c]imidazol-2-yl)-2-methylbenzofuran-7-carbonitrile

10A. 2-Allyloxy-4-nitrobenzonitrile

To a solution of 2-hydroxy-4-nitro-benzonitrile [164 mg, 1.0 mmol,prepared as described in Yasubiro Imakura et. al. Chem. Pharm. Bull. 40(7), 1691-1696 (1992)] in anhydrous DMF (2 mL) was added allyl bromide(0.11 mL, 1.3 mmol), followed by K₂CO₃ (166 mg, 1.2 mmol). The resultingsuspension was heated to 50° C. under Argon for 4 h. After cooling tort, the reaction mixture was poured into ice/water, and extracted withEtOAc (3×). The combined EtOAc extracts were washed with brine, dried(Na₂SO₄), and concentrated under reduced pressure. The crude product waschromatographed (silica gel) eluting with 30% to 60% EtOAc/hexane tofurnish the title compound (160 mg, 80% yield) as a light yellow solid.

10B. 2-Allyloxy-4-aminobenzonitrile

The title compound (1.70 g) as a light yellow solid was prepared fromcompound 10A (2.04 g, 10 mmol) by procedures analogous to thosedescribed in Experiment 6C.

10C. N-(3-Allyloxy-4-cyanophenyl)-2,2-dimethylpropionamide

To a solution of 10B (1.9 g, 10.9 mmol) in anhydrous CH₂Cl₂ (16 mL)cooled at 0° C. was added 1 N aqueous NaOH solution (16.4 mL, 16.4mmol), followed by pivaloyl chloride (1.9 mL, 15.3 mmol). After theaddition, the reaction mixture was stirred at 0° C. for 2 h, thenconcentrated under reduced pressure to remove most of the CH₂Cl₂solvent. The remaining residue was diluted with water, and the resultingprecipitate collected by filtration, washed with water, hexane, anddried under vacuum to furnish the title compound as a light brown solid(2.6 g).

10D. N-(2-Allyl-4-cyano-3-hydroxyphenyl)-2,2-dimethylpropionamide

A solution of compound 10C (1.5 g, 5.8 mmol) in NMP (7 mL) was heated to220° C. for 3 h. After cooling to rt, the reaction mixture was pouredinto ice/water, and extracted with EtOAc (3×). The combined EtOAcextracts were washed with brine, dried (Na₂SO₄), and concentrated underreduced pressure. The crude product was chromatographed (silica gel)eluting with 0% to 50% EtOAc/hexane to furnish the title compound (0.7g).

10E. N-(7-Cyano-2-methyl-benzofuran-4-yl)-2,2-dimethylpropionamide

To a solution of compound 10D (0.5 g, 1.94 mmol) in a mixed solvent ofDMF (2.5 mL) and water (1.9 mL) was added Cu(OAc)₂ (1.057 g, 5.82 mmol),followed by a 10 M aqueous solution of LiCl (0.58 mL, 5.8 mmol) andPdCl₂ (34.3 mg, 0.194 mmol). The resulting suspension was heated to 100°C. for 1 h. After cooling to rt, the reaction mixture was poured intoice/water, and extracted with EtOAc (3×). The combined EtOAc extractswere washed with brine, dried (Na₂SO₄), and concentrated under reducedpressure. The crude product was chromatographed (silica gel) elutingwith 10% to 50% EtOAc/hexane to furnish the title compound (0.3 g) as awhite foam.

10F.(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-methylbenzofuran-7-carbonitrile

The title compound was prepared from compound 10E by proceduresanalogous to those described in Experiment 2 (2D to 2F). HPLC: 99% at4.2 min (retention time) (Conditions: Zorbax SB C18 (4.6×75 mm); Elutedwith 0% to 100% B, 8 min gradient. (A=90% H₂O—10% MeOH—0.1% H₃PO₄ andB=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5 mL/min. UV detection at220 nm). Chiral HPLC: retention time=10.99 min (99%); Conditions: OD(4.6×250 mm); Eluted with 25% isopropanol in hexane for 30 min at 1mL/min. MS (ES) m/z 312 [M+1]⁺.

EXAMPLE 11(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-methylbenzoxazole-7-carbonitrile

11A. 3-Amino-2-hydroxy-4-nitro-benzonitrile

To a solution of 2-hydroxy-4-nitro-benzonitrile [1.0 g, 6.1 mmol,prepared as described in Yasubiro Imakura et. al. Chem. Pharm. Bull. 40(7), 1691-1696 (1992)] in DMSO (60 mL) was added trimethylhydraziniumiodide (1.23 g, 6.09 mmol), followed by sodium tert-pentoxide (2.01 g,6.09 mmol). The mixture was stirred at rt overnight, and thenpartitioned between 10% HCl and ether. The separated ether layer waswashed with H₂O, brine, dried (Na₂SO₄), and concentrated under reducedpressure to furnish the title compound (0.78 g) as a brown solid.

11B. 2-Methyl-4-nitro-benzoxazole-7-carbonitrile

To a solution of 11A (0.2 g, 1.12 mmol) in xylene (6 mL) was addedtriethyl orthoacetate (0.62 mL, 3.35 mmol) and pyridiniump-toluenesulfonate (0.04 g, 1.12 mmol). The mixture was refluxed for 3h, then allowed to cool to rt and partitioned between H₂O and EtOAc. Theseparated EtOAc layer was concentrated under reduced pressure and theresidue was chromatographed eluting with 20% EtOAc in hexane to affordcompound 11B (0.16 g) as a solid.

11C. 4-Amino-2-methylbenzoxazole-7-carbonitrile

To a solution of 11B (0.16 g, 0.79 mmol) in EtOAc (2 mL) was added ironpowder (0.1 g), followed by 10% aqueous AcOH (2 mL). The mixture wasstirred at 60° C. for 2 h, then allowed to cool to rt and partitionedbetween saturated NaHCO₃ and EtOAc. The separated EtOAc layer wasconcentrated under reduced pressure to give the title compound (0.14 g)as a solid.

11D.(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydro-pyrrolo[1,2-c]imidazol-2-yl)-2-methylbenzoxazole-7-carbonitrile

The title compound was prepared from 11C by procedures analogous tothose described in Example 2 (2E to 2F) to give the product as a whitefoam. HPLC: 99% pure at 10.55 min (retention time) (CHIRALPAK® OD column4.6×250 mm; 25% isopropanol in hexane over 30 min, 1 mL/min, monitoringat 220 nm); MS (ES) m/z 313 [M+1]⁺.

EXAMPLE 12(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)quinoline-8-carbonitrile

12A. 8-Methyl-5-nitroquinoline

To a solution of commercially available 8-methylquinoline (5.0 g, 34mmol) in concentrated H₂SO₄ (19 mL) at 0° C. was added portionwise KNO₃(4.29 g, 42.4 mmol). After the addition, the reaction was stirred at rtfor 17 h, then poured into ice/water and extracted with EtOAc (3×100mL). The aqueous layer was basified to pH=9 with solid Na₂CO₃ andextracted with EtOAc (2×100 mL). The combined extracts were washed withbrine, dried (Na₂SO₄), filtered and concentrated under reduced pressure.The residue was chromatographed (silica gel) to yield the title compound(5.98 g, 94%) as a light yellow solid. LC/MS m/z 189 [M+H]⁺.

12B. 5-Nitroquinoline-8-carboxylic acid

Compound 12A (1.0 g, 5.3 mmol) was dissolved in concentrated H₂SO₄ (7mL), cooled to 0° C., and then CrO₃ (2.12 g, 21.3 mmol) was addedportionwise over 30 min. After the addition, the reaction mixture waswarmed to rt and then heated to 80° C. for 1 h. After cooling to rt, thereaction was diluted with water, basified with 15% aqueous NaOH, thenreacidified to pH=3 with AcOH, and extracted with EtOAc (4×100 mL). Thecombined EtOAc extracts were washed with brine, dried (Na₂SO₄), filteredand concentrated under reduced pressure to yield the title compound (760mg, 66%) as a yellow solid. LC/MS m/z 219 [M+H]⁺.

12C. 5-Nitroquinoline-8-carbonitrile

To a suspension of compound 12B (600 mg, 2.75 mmol) in anhydrous THF (25mL) cooled to −15° C. was added Et₃N (0.46 mL, 3.3 mmol), followed by adropwise addition of ethyl chloroformate (0.33 mL, 3.44 mmol). Thereaction mixture was stirred at −15° C. for 30 min, then NH₃ gas wasbubbled into the reaction for 5 min followed by warming of the reactionto rt for 1h. The solvent was evaporated to give 850 mg (>100%) of theamide as a yellow solid which was carried on to the next step withoutfurther purification. The amide (850 mg) was dissolved in pyridine (25mL), and imidazole (377 mg, 5.49 mmol) was added. The mixture was cooledto −30° C. under nitrogen, POCl₃ (1.01 mL, 10.7 mmol) was added and thereaction was warmed to 0° C. for 30 min, and then evaporated to dryness.The residue was chromatographed (silica gel) eluting with CH₂Cl₂ toafford the title compound (416 mg, 76%, 2 steps). LC/MS m/z 200 [M+H]⁺.

12D.(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-quinoline-8-carbonitrile

The title compound was synthesized from compound 12C by proceduresanalogous to those described in Experiment 3A and Experiment 2 (2E to2F). 99.8% at 2.05 min (retention time) (Conditions: YMC S5 ODS (4.6×50mm); Eluted with 0% to 100% B; 4 min gradient; (A=90% H₂O—10% MeOH—0.1%H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 4 mL/min. UVdetection at 220 nm). LC/MS m/z 309 [M+H]⁺.

EXAMPLE 13(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)isoquinoline-1-carbonitrile

13A. 4-Bromoisoquinoline 2-oxide

A solution of 4-bromoisoquinoline (4.16 g, 18.6 mmol) in chloroform (100mL) was added dropwise over 1 h to a solution of 70% mCPBA (12.4 g, 50.3mmol) in chloroform (100 mL) at rt. After stirring 18 h, the reactionmixture was washed with 1 N NaOH (2×150 mL), dried (MgSO₄) andconcentrated under reduced pressure to afford compound 13A (4.23 g, 94%)as an off-white solid. ¹H NMR(400 MHz, CDCl₃) □ 8.71 (s, 1H), 8.43 (s,1H), 8.09 (d, 1H, J=8 Hz), 7.70 (m, 3H).

13B. 4-Bromoisoquinoline-1-carbonitrile

DBU (1.67 mL, 11.2 mmol) was added to a mixture of compound 13A (1.12 g,5.00 mmol) and cyanotrimethylsilane (0.75 mL, 5.5 mmol) in THF (35 mL).The resulting homogeneous mixture was refluxed for 20 min. Afterconcentrating under reduced pressure, the residue was purified by flashchromatography on a 5×15 cm silica gel column, eluting with 3:1hexane:EtOAc to give compound 13B (0.95 g, 82%) as a white powder. ¹HNMR (400 MHz, CDCl₃) □ 8.85 (s, 1H), 8.36 (d, 1H, J=8.5 Hz), 8.28 (d,1H, J=8.5 Hz), 7.96 (t, 1H, J=8.5 Hz), 7.89 (t, 1H, J=8.5 Hz).

13C. 4-(2,4-Dimethoxybenzylamino)isoquinoline-1-carbonitrile

A mixture of compound 13B (699 mg, 3.00 mmol) and2,4-dimethoxybenzylamine (4.8 mL, 30 mmol) in acetonitrile (15 mL) wasrefluxed for 16 h. After concentration under reduced pressure, theresidue was purified on a 5×15 cm silica gel column, eluting with 3:2hexane:EtOAc to afford compound 13C (290 mg, 30%) as a light yellowsolid. HPLC: 1.76 min (retention time) (Phenomenex C-18, 5 microncolumn, 4.6×30 mm, eluting with 10-90% aqueous MeOH over 2 mincontaining 0.1% TFA, 4 mL/min, monitoring at 254 nm).

13D. 4-Aminoisoquinoline-1-carbonitrile

Compound 13C (50 mg, 0.16 mmol) was treated with TFA (0.5 mL) for 1 h.The highly colored mixture was partitioned between EtOAc (30 mL) and 1 NNaOH (30 mL). After washing with brine (15 mL), the organic layer wasdried (MgSO₄) and concentrated under reduced pressure to afford compound13D (24 mg, 92%) as a yellow solid. HPLC: 99% at 1.09 min (retentiontime) (Phenomenex C-18, 5 micron column, 4.6×30 mm, eluting with 10-90%aqueous MeOH over 2 min containing 0.1% TFA, 4 mL/min, monitoring at 254nm). MS (ES+) m/z 170 [M+H]⁺.

13E.(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)isoquinoline-1-carbonitrile

The title compound was synthesized from compound 13D by proceduresanalogous to those described in Experiment 2E to 2F. HPLC: 97.6% at 1.54min (retention time) (Conditions: YMC S5 C-18 (4.6×50 mm); Eluted with0% to 100% B; 4 min gradient; (A=90% H₂O—10% MeCN—0.1% TFA and B=10%H₂O—90% MeCN—0.1% TFA); Flow rate at 4 mL/min. UV detection at 220 nm).LC/MS m/z 309 [M+H]⁺.

EXAMPLE 14(7R,7aS)-3-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-methoxybenzonitrile

14A. 3-Amino-2-methoxy-4-nitrobenzonitrile

To a stirring suspension of 11A (0.34 g, 1.86 mmol) and K₂CO₃ (0.283 g,2.05 mmol) in DMF (3 mL) was added iodomethane (0.127 mL, 2.05 mmol).After the addition, the reaction mixture was stirred at rt for 16 h,then partitioned between H₂O and CH₂Cl₂. The separated CH₂Cl₂ layer waswashed with water, brine, dried (Na₂SO₄), filtered and concentratedunder reduced pressure. The residue was chromatographed (silica gel)eluting with 5% to 40% EtOAc/hexane to yield the title compound (0.22 g)as a solid.

14B. 3-Chloro-2-methoxy-4-nitrobenzonitrile

To a suspension of CuCl₂ (0.11 g, 0.78 mmol) in CH₃CN (2 mL) at rt wasadded tert-butyl nitrite (0.1 mL, 0.84 mmol). After addition, themixture was heated to 65° C. while a solution of 14A (0.125 g, 0.647mmol) in CH₃CN (3 mL) was slowly added. After stirring at 65° C. for 1h, the mixture was allowed to cool to rt, and then partitioned betweenH₂O and CH₂Cl₂. The CH₂Cl₂ layer was washed with water, brine, dried(Na₂SO₄), filtered and concentrated under reduced pressure. The residuewas chromatographed (silica gel) eluting with 5% to 60% EtOAc/hexane toyield the title compound (0.085 g).

14C. 4-Amino-3-chloro-2-methoxybenzonitrile

The title compound was prepared from compound 14B in a manner similar tothat described in Experiment 11C.

14D.(7R,7aS)-3-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-methoxybenzonitrile

The title compound was prepared from 14C by procedures analogous tothose described in Example 2E to 2F. HPLC: 99% pure at 13.64 min(retention time) (CHIRALPAK® OD column 4.6×250 mm; 25% isopropanol inhexane over 30 min, 1 mL/min, monitoring at 220 nm); MS (ES) m/z 316[M+1]⁺.

EXAMPLE 15(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-methoxy-3-methylbenzonitrile

The title compound was prepared from commercially available2-methyl-3-nitroanisole by procedures analogous to those described inExample 3A and Example 2. HPLC: 100% at 3.63 min (retention time)(Conditions: Zorbax SB C18 (4.6×75 mm); Eluted with 0% to 100% B, 8 mingradient. (A=90% H₂O—10% MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1%H₃PO₄); Flow rate at 2.5 mL/min. UV detection at 220 nm). Chiral HPLC:retention time=9.76 min (99%); Conditions: OD (4.6×250 mm); Eluted with25% isopropanol in hexane for 30 min at 1 mL/min. MS (ES) m/z 302[M+1]⁺.

EXAMPLE 16(7R,7aS)-2-Hydroxy-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To compound 14 (90 mg, 0.3 mmol) in a 5-mL round bottom flask at 0° C.was added a 1.0 M solution of BBr₃ in CH₂Cl₂ (1 mL, 1 mmol). Thereaction mixture was stirred at 0° C. for 30 min, then at rt foradditional 30 min. After cooling at 0° C., MeOH (3 mL) was added, thereaction was stirred at 0° C. for 30 min, and then concentrated underreduced pressure to obtain a crude product, which was chromatographed(silica gel) eluting with 0% to 5% MeOH in CH₂Cl₂ to yield the titlecompound (12 mg). HPLC: 98% at 2.83 min (retention time) (Conditions:Zorbax SB C18 (4.6×75 mm); Eluted with 0% to 100% B, 8 min gradient.(A=90% H₂O—10% MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flowrate at 2.5 mL/min. UV detection at 220 nm). Chiral HPLC: retentiontime=16.13 min (99%); Conditions: OD (4.6×250 mm); Eluted with 25%isopropanol in hexane for 30 min at 1 mL/min. MS (ES) m/z 288 [M+1]⁺.

EXAMPLE 17(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methoxy-2-(trifluoromethyl)benzonitrile

17A. 2-Nitro-6-(trifluoromethyl)phenol

To a solution of commercially available 2-trifluoromethylphenol (2.5 g,15.42 mmol) in AcOH (3 mL) cooled to 0° C. was added dropwiseconcentrated HNO₃ (1.5 mL). After the addition, the mixture was stirredat at 0° C. for 5 min, at rt for 10 min, then poured into ice/water, andextracted with ether (3×). The combined ether extracts were washed withwater, brine, dried (Na₂SO₄) and concentrated under reduced pressure.The residue was chromatographed (silica gel) eluting with 0% to 10%EtOAc/hexane to yield the title compound (1.4 g, 44% yield).

17B. 2-Nitro-6-(trifluoromethyl)anisole

To a suspension of compound 17A (1.4 g, 6.76 mmol) and K₂CO₃ (1.4 g, 10mmol) in DMF (60 mL) was added iodomethane (0.46 mL, 7.44 mmol). Afterthe addition, the mixture was stirred at 40° C. overnight, then cooledto rt, partitioned between ether and water. The separated ether layerwas washed with water and brine, then dried (Na₂SO₄) and concentratedunder reduced pressure. The residue was chromatographed (silica gel)eluting with 10% EtOAc/hexane to yield the title compound (1.38 g, 92%yield).

17C. 2-Methoxy-3-(trifluoromethyl)aniline

The title compound (980 mg) was prepared from compound 17B in a mannersimilar to that described in Experiment 11C.

17D. 4-Bromo-2-methoxy-3-(trifluoromethyl)aniline

To a solution of compound 17C (200 mg, 1.05 mmol) in CH₂Cl₂ (2 mL)cooled to −20° C. was added dropwise a solution of2,4,4,6-tetrabromo-2,5-cyclohexadienone (429 mg, 1.05 mmol) in CH₂Cl₂ (2mL). After the addition, the reaction was stirred at rt overnight, thenadditional amount of 2,4,4,6-tetrabromo-2,5-cyclohexadienone (429 mg,1.05 mmol) was added. The reaction was continued for 2 more days, thenconcentrated under reduced pressure. The residue was chromatographed(silica gel) eluting with 10%-20% EtOAc/hexane to yield the titlecompound (47mg, 16% yield).

17E. N-(4-Bromo-2-methoxy-3-(trifluoromethylphenyl)acetamide

The title compound (237 mg) was prepared from compound 17D in a mannersimilar to that described in Experiment 2A.

17F.(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methoxy-2-trifluoromethylbenzonitrile

The title compound (55 mg) was prepared from 17E by procedures analogousto those described in Example 2 (2C to 2F). HPLC: 99% at 2.25 min(retention time) (Conditions: Phenom. Lura C18 (4.6×50 mm); Eluted with0% to 100% B, 4 min gradient (A=90% H₂O—10% MeOH—0.1% TFA and B=10%H₂O—90% MeOH—0.1% TFA); Flow rate at 4.0 mL/min. UV detection at 220nm.). Chiral HPLC: 99% pure at 8.98 min (retention time) (CHIRALPAK® ODcolumn 4.6×250 mm; 30% isopropanol in hexane over 30 min, 1 mL/min,monitoring at 220 nm); MS (ES) m/z 356 [M+1]⁺.

EXAMPLE 18(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methyl-2-trifluoromethylbenzonitrile

18A. N-(4-Chloro-3-trifluoromethylphenyl)-2,2-dimethylpropionamide

To a solution of commercially available4-chloro-3-(trifluoromethyl)aniline (15.0 g, 76.7 mmol) in anhydrous THF(200 mL) cooled to 0-5° C. was added triethylamine (11.7 mL, 84.4 mmol)followed by pivaloyl chloride (10.4 mL, 84.4 mmol) over 30 min. The icebath was removed and the mixture stirred at rt for 1 h. The mixture wasdiluted with ether and filtered. The filtrate was washed with water (2×)and brine, dried (MgSO₄), filtered and concentrated. The residue wastriturated with hexanes and the solid was filtered and dried undervacuum to afford compound 18A (20.4 g, 95%); MS (ES) m/z 280 [M+1]⁺.

18B.N-(4-Chloro-2-methyl-3-trifluoromethylphenyl)-2,2-dimethyl-propionamide

To a solution of compound 18A (2.29 g, 8.19 mmol) in anhydrous THF (25mL) cooled to 0-5° C. was added a solution of 1.6 M n-butyllithium inhexanes (12.3 mL, 19.7 mmol) slowly, so that the reaction temperaturewas maintained below 5° C. The solution was stirred at 0-5° C. for 1.5h. A solution of iodomethane (0.56 mL, 9.01 mmol) in petroleum ether (2mL) was then added over 20 min while maintaining the temperature below5° C. The suspension was stirred at 0-5° C. for 1 h and diluted withwater and ether. The aqueous layer was extracted with ether and thecombined organic layers washed with brine, dried (MgSO₄), filtered andconcentrated. The residue was chromatographed (silica gel), eluting withCH₂Cl₂ to afford the title compound (1.60 g, 67%). MS (ES) m/z 294[M+1]⁺.

18C.N-(4-Cyano-2-methyl-3-trifluoromethylphenyl)-2,2-dimethyl-propionamide

A suspension of compound 18B (8.36 g, 28.5 mmol) and CuCN (4.33 g, 65.5mmol) in anhydrous N-methylpyrrolidinone (85 mL) was refluxed for 38 h.After cooling to rt, the suspension was poured into ice/water withstirring. The solid was filtered, washed with water and dried to yieldan 85:15 mixture (7.55 g) of compounds 18C and 18D.

18D. 4-Amino-3-methyl-2-trifluoromethylbenzonitrile

A solution of the mixed product from 18C (7.53 g, 26.5 mmol) in 120 mLof concentrated HCl/EtOH (1:1) was refluxed for 14 h. After cooling tort, the solution was concentrated under reduced pressure. The resultingresidue was dissolved in EtOAc, washed with saturated aqueous NaHCO₃(2×) and brine (1×), dried (MgSO₄), filtered and concentrated. Theresidue was chromatographed (silica gel), eluting with chloroform/MeOH(98:2) to furnish the title compound (4.62 g, 87%). MS (ES) m/z 201[M+1]⁺.

18E. 4-Isocyanato-3-methyl-2-trifluoromethylbenzonitrile

The title compound was prepared from 18D in a manner similar to thatdescribed in Experiment 2E.

18F.(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methyl-2-trifluoromethylbenzonitrile

The title compound was prepared (3.0 g, 53% yield) by proceduresanalogous to those described in Experiment 2 F. HPLC: 98% at 2.33, 2.58min (retention time) (Conditions: Phenom. Lura C18 (4.6×50 mm); Elutedwith 0% to 100% B, 4 min gradient (A=90% H₂O—10% MeOH—0.1% TFA and B=10%H₂O—90% MeOH—0.1% TFA); Flow rate at 4.0 mL/min. UV detection at 220nm.). Chiral HPLC: retention time=9.71 min (98%); Conditions: OD(4.6×250 mm); Eluted with 25% isopropanol in hexane for 30 min at 1mL/min. MS (ES) m/z 340 [M+1]⁺.

EXAMPLE 19(7S,7aR)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methyl-2-trifluoromethylbenzonitrile

To a stirring solution of compound 1A (182 mg, 1 mmol) in MeOH (10 mL)was added WA21J resin (1 g) in one portion. The resulting suspension wasstirred at rt for 30 min, then filtered and the filtrate concentratedunder reduced pressure to give the corresponding free amine as acolorless oil. To a solution of the amine in CH₂Cl₂ (2 mL) was added asolution of 18E (0.7 mmol) in CH₂Cl₂ (2 mL), followed by 4 Å molecularsieves (0.5 g). The resulting suspension was stirred at rt for 20 h, andthen DBU (0.2 mL, 1.5 mmol) was added. After stirring at rt for 3 days,the reaction mixture was filtered and the filtrate washed with 1 Naqueous HCl, water, brine, dried (MgSO₄), filtered and concentrated. Theresidue was chromatographed (silica gel), eluting with 5% MeOH inEtOAc/hexane (1:1) to afford the title compound (45 mg) as a whitesolid. HPLC: 100% at 4.32, 4.89 min (retention time) (Conditions: ZorbaxSB C18 (4.6×75 mm); Eluted with 0% to 100% B, 8 min gradient (A=90%H₂O—10% MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at2.5 mL/min. UV detection at 220 nm.). Chiral HPLC: retention time=14.11min (99%); Conditions: OD (4.6×250 mm); Eluted with 20% isopropanol inhexane for 30 min at 1 mL/min. MS (ES) m/z 340 [M+1]⁺.

EXAMPLE 20(7R,7aS)-2-Bromo-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

20A. 4-Amino-2-bromo-3-methylbenzaldehyde

To a solution of commercially available 3-bromo-2-methyl-phenylamine(1.86 g, 10 mmol) in DMSO (200 mL) was added concentrated HCl (10 mL),followed by CuCl₂ (2.7 g, 20 mmol). The resulting suspension was heatedto 90° C. for 6 h. After cooling to rt, the reaction mixture was pouredinto ice/water (600 mL) and adjusted to pH=8 by dropwise addition of 10%aqueous NaOH. The resulting greenish suspension was filtered through apad of Celite®, and the filtrate extracted with ether (3×). The combinedorganics were washed with brine, dried (Na₂SO₄), filtered andconcentrated. The residue was chromatographed (silica gel), eluting with30-70% EtOAc/hexane to furnish the title compound (0.67 g, 31%).

20B. 4-Amino-2-bromo-3-methylbenzonitrile

To a solution of hydroxylamine hydrochloride (109 mg, 1.58 mmol) in H₂O(0.37 mL) was added compound 18A (321 mg, 1.5 mmol), followed bypyridine (0.75 mL). The reaction was stirred at rt for 1 h, then CuSO₄(75 mg, 0.3 mmol) was added, followed by Et₃N (0.44 mL, 3.2 mmol) and asolution of DCC (371 mg, 1.8 mmol) in CH₂Cl₂ (3 mL). After addition, thereaction mixture was stirred at rt until the oxime was consumed (1 h).The reaction was then treated with formic acid (0.26 mL) and stirred atrt for another 10 min to consume the excess DCC. The reaction wasfiltered through a pad of Celite®, and the filtrate partitioned betweenCH₂Cl₂ and saturated aqueous NaHCO₃. The separated organic layer waswashed with brine, dried (Na₂SO₄), filtered and concentrated. Theresidue was chromatographed (silica gel), eluting with 10-50%EtOAc/hexane to furnish the title compound (260 mg, 83%).

20C.(7R,7aS)-2-Bromo-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

The title compound was prepared from 18B by procedures analogous tothose described in Experiment 2E and 2F. HPLC: 99% at 3.99, 4.53 min(retention time) (Conditions: Zorbax SB C18 (4.6×75 mm); Eluted with 0%to 100% B, 8 min gradient (A=90% H₂O—10% MeOH—0.1% H₃PO₄ and B=10%H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5 mL/min. UV detection at 220nm). Chiral HPLC: retention time=15.4 min (99%); Conditions: OD (4.6×250mm); Eluted with 20% isopropanol in hexane for 30 min at 1 mL/min. MS(ES) m/z 351 [M+1]⁺.

EXAMPLE 21(7R,7aS)-2-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3,6-dimethylbenzonitrile

21A. 1-Chloro-2,5-dimethyl-3-nitrobenzene

To a solution of commercially available 2-chloro-1,4-dimethylbenzene (5mL, 37 mmol) in concentrated H₂SO₄ (5 mL) cooled to 0-5° C. was addeddropwise concentrated HNO₃ (4.7 mL, 74.6 mmol) over 20 min. After theaddition, the reaction was stirred at 0-5° C. for 30 min, then pouredcarefully into a mixture of ice and saturated aqueous K₂CO₃ solution (40mL), and extracted with EtOAc (3×). The combined EtOAc extracts werewashed with brine, dried (Na₂SO₄), filtered and concentrated underreduced pressure. The residue was chromatographed (silica gel), elutingwith 10-30% EtOAc/hexane to give a mixture of the title compound and itsregioisomer (1-chloro-2,5-dimethyl-4-nitrobenzene) (3.0 g). The mixturewas further chromatographed (silica gel) eluting with 10% CH₂Cl₂ inhexane to afford pure compound 19A (0.2 g).

21B. 3-Chloro-2,5-dimethylaniline

The title compound (1.70 g) was prepared from 21A (2.15 g, 11.6 mmol) ina manner similar to that described in Experiment 6C.

21C. 4-Bromo-3-chloro-2,5-dimethylaniline

To a solution of compound 21B (1.56 g, 10 mmol) in CHCl₃ (30 mL) cooledto 0-5° C. was added portionwise tetrabutylammonium tribromide (434 mg,9.0 mmol). After addition, the reaction was stirred at 0-5° C. for 5min, then quenched with saturated aqueous NaHCO₃ (30 mL) and 5% aqueousNa₂S₂O₃ (30 mL). The mixture was stirred at rt for 10 min, thenextracted with CH₂Cl₂ (3×). The combined organics were washed withbrine, dried (Na₂SO₄), filtered and concentrated. The residue waschromatographed (silica gel), eluting with 20-60% EtOAc/hexane to givethe title compound (0.75 g, 32% yield).

21D. N-(4-Bromo-3-chloro-2,5-dimethylphenyl)acetamide

The title compound (0.82 g) was prepared from 21C (0.74 g, 3.17 mmol) ina manner similar to that described in Experiment 2A.

21E.(7R,7aS)-2-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2c]imidazol-2-yl)-3,6-dimethylbenzonitrile

The title compound (30 mg) as a white solid was prepared by proceduresanalogous to those described in Experiment 2C to 2F. HPLC: 99% at 4.50,4.87 min (retention time) (Conditions: Zorbax SB C18 (4.6×75 mm); Elutedwith 0% to 100% B, 8 min gradient. (A=90% H₂O—10% MeOH—0.1% H₃PO₄ andB=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5 mL/min. UV detection at220 nm). Chiral HPLC: retention time=12.07 min (99%); Conditions: OD(4.6×250 mm); Eluted with 20% isopropanol in hexane for 30 min at 1mL/min. MS (ES) m/z 320 [M+1]⁺.

EXAMPLE 22(7R,7aS)-2-(4-Bromo-3-chloro-2-methylphenyl)-7-hydroxytetrahydropyrrolo[1,2-c]imidazole-1,3-dione

22A. N-(4-Bromo-3-chloro-2-methylphenyl)acetamide

The title compound was prepared (2.75 g, 96% yield) from commerciallyavailable 3-chloro-2-methylaniline by procedures analogous to thosedescribed in Experiment 2A and 2B.

22B. 4-Bromo-3-chloro-2-methylaniline

The title compound was prepared (0.70 g, 83% yield) from 22A in a mannersimilar to that described in Experiment 2D.

22C.(7R,7aS)-2-(4-Bromo-3-chloro-2-methylphenyl)-7-hydroxytetrahydropyrrolo[1,2-c]imidazole-1,3-dione

The title compound was prepared (0.71 g, 88% yield) by proceduresanalogous to those described in Experiment 2E and 2F. HPLC: 99% at 2.78,2.95 min (retention time) (Conditions: Phenom. Lura C18 (4.6×50 mm);Eluted with 0% to 100% B, 4 min gradient (A=90% H₂O—10% MeOH—0.1% TFAand B=10% H₂O—90% MeOH—0.1% TFA); Flow rate at 4.0 mL/min. UV detectionat 220 nm). Chiral HPLC: retention time=7.17 min (92%); Conditions: OD(4.6×250 mm); Eluted with 25% isopropanol in hexane for 30 min at 1mL/min MS (ES) m/z 360 [M+1]⁺.

EXAMPLE 23(7R,7aS)-2-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

23A. 3-Chloro-2-methylphenylacetamide

To a solution of 3-chloro-2-methylaniline (3.00 g, 21.2 mmol) in 25 mLof EtOH at rt was added acetic anhydride (2.40 mL, 25.4 mmol), and thesolution was stirred at rt for 2 h. The mixture was concentrated underreduced pressure to give 3.89 g (100%) of the desired acetamide. ¹H NMR(DMSO-d₆) δ 2.05 (s, 3H), 2.20 (s, 3H), 7.16 (t, J=7.7, 8.3, 1H), 7.25(d, J=8.3, 1H), 7.31 (d, J=8.3, 1H), 9.55 (s, 1H); ¹³C NMR (DMSO-d₆) δ15.1, 23.1, 124.4, 125.8, 126.7, 130.3, 133.7, 138.0, 168.3; HPLC a)column: Phenominex ODS C18 4.6×50 mm, 4 min gradient, 10% MeOH/90%H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA; 1 min hold, 4 mL/min UVdetection at 220 nm, 2.32 min retention time; HPLC b) column: ShimadzuShim-Pack VP-ODS C18 4.6×50 mm, 4 min gradient, 10% MeOH/90% H₂O/0.1%TFA to 90% MeOH/10% H₂O/0.1% TFA, 1 min hold; 4 mL/min, UV detection at220 nm, 2.20 min retention time (99%); MS (ES) m/z 184 [M+H]⁺.

23B. 4-Bromo-3-chloro-2-methylphenylacetamide

To a suspension of acetamide 23A (2.00 g, 10.9 mmol) in 15 mL of glacialAcOH cooled to approximately 15° C. was added bromine (1.67 mL, 32.7mmol) over 20 min. The ice bath was removed and the solution was stirredfor 2 h, poured into ice water with stirring, and the solid was thenfiltered and dried to give 2.75 g (96%) of the desired bromide. ¹H NMR(DMSO-d₆) δ 2.05 (s, 3H), 2.28 (s, 3H), 7.29 (d, J=8.3, 1H), 7.56 (d,J=8.8, 1H), 9.60 (s, 1H); ¹³C NMR (DMSO-d₆) δ 16.7, 23.1, 118.1, 125.5,130.4, 132.7, 133.4, 137.1, 168.4; HPLC a) column: Phenominex ODS C184.6×50 mm, 4 min gradient, 10% MeOH/90% H₂O/0.1% TFA to 90% MeOH/10%H₂O/0.1% TFA, 1 min hold, 4 mL/min, UV detection at 220 nm, 2.95 minretention time; HPLC b) column: Shimadzu Shim-Pack VP-ODS C18 4.6×50 mm,4 min gradient, 10% MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA,1 min hold, 4 mL/min, UV detection at 220 nm, 2.87 min retention time(98%); MS (ES) m/z 263 [M+H]⁺.

23C. 3-Chloro-4-cyano-2-methylphenylacetamide

A suspension of bromide 23B (2.70 g, 10.3 mmol) and copper cyanide (0.92g, 10.3 mmol) in DMF (30 mL) was heated to 150° C. for 4 h. Thesuspension was cooled, poured into water with stirring, and the solidwas filtered and dried to give 1.44 g (67%) of the desired nitrile. ¹HNMR (DMSO-d₆) δ 2.12 (s, 3H), 2.29 (s, 3H), 7.72 (d, J=8.8, 1H), 7.75(d, J=8.2, 1H), 9.73 (s, 1H); ¹³C NMR (DMSO-d₆) δ 15.3, 23.5, 107.7,116.5, 123.0, 130.1, 131.5, 135.7, 142.3, 168.8; HPLC a) column:Phenominex ODS C18 4.6×50 mm, 4 min gradient, 10% MeOH/90% H₂O/0.1% TFAto 90% MeOH/10% H₂O/0.1% TFA, 1 min hold, 4 mL/min, UV detection at 220nm, 2.23 min retention time; HPLC b) column: Shimadzu Shim-Pack VP-ODSC18 4.6×50 mm, 4 min gradient, 10% MeOH/90% H₂O/0.1% TFA to 90% MeOH/10%H₂O/0.1% TFA, 1 min hold, 4 mL/min, UV detection at 220 nm, 2.13 minretention time (95%); MS (ES) m/z 209 [M+H]⁺.

23D. 3-Chloro-4-cyano-2-methylphenylaniline

A solution of cyanoacetamide 23C (9.90 g, 47.4 mmol) in 100 mL ofconcentrated HCl/EtOH (1:1) was refluxed 30 min. The solution was thenconcentrated and dried under reduced pressure to give 9.41 g (98%) ofthe desired aniline as the hydrochloride salt. The free base of theaniline was obtained by suspending the salt in EtOAc and washing withsaturated aqueous NaHCO₃ solution. The organic layer was then dried(MgSO₄), filtered and concentrated under reduced pressure. ¹H NMR(DMSO-d₆) δ 2.12 (s, 3H), 6.30 (s, 2H), 6.61 (d, J=8.23, 1H), 7.36 (d,J=8.23, 1H); ¹³C NMR (DMSO-d₆) δ 13.8, 96.9, 112.1, 118.3, 118.85,132.2, 135.6, 152.5; HPLC a) column: Phenominex ODS C18 4.6×50 mm, 4 mingradient, 10% MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA, 1 minhold, 4 mL/min, UV detection at 220 nm, 2.43 min retention time; HPLCb):column: Shimadzu Shim-Pack VP-ODS C18 4.6×50 mm, 4 min gradient, 10%MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA, 1 min hold, 4mL/min, UV detection at 220 nm, 2.31 min retention time (99%); MS (ES)m/z 167 [M+H]⁺.

23E. 2-Chloro-4-isocyanato-3-methylbenzonitrile

The title compound was prepared from compound 23D in a manner similar tothat described in Experiments 2D to 2E.

23F.(2S,3R)-1-(3-Chloro-4-cyano-2-methylphenylcarbamoyl)-3-hydroxypyrrolidine-2-carboxylicacid methyl ester

To a solution of hydroxyproline compound 1F (493 mg, 3.40 mmol) inCH₂Cl₂ (15 mL) was added 4 Å molecular sieves (˜3.0 g), followed byisocyanate 23E (725 mg, 3.22 mmol), and the resulting mixture wasstirred at rt overnight, filtered, and concentrated under reducedpressure. The residue was purified by flash chromatography (silica gel,0.5% MeOH in EtOAc/hexane, 1:1) to afford the title compound (736 mg) asan off-white solid. HPLC column: YMC S-5 C18 (4.6×50 mm), 0% to 100% B,4 min gradient, 1 min hold (A=90% H₂O—10% CH₃CN—0.1% TFA and B=10%H₂O—90% CH₃CN—0.1% TFA), flow rate at 4 mL/min, UV detection at 220 nm,1.57 min retention time (100%); MS (ES) m/z 338 [M+H]⁺.

23G.(7R,7aS)-2-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile.

To a suspension of cis-3-hydroxyproline methyl ester, HCl salt (4.91 g,27 mmol) in CH₂Cl₂ (100 mL) cooled to 0° C. was added i-Pr₂NEt (4.79 mL,27.5 mmol). After stirring at rt for 15 min, isocyanate 23E was added asa solid in one portion through a powder addition funnel, rinsing with 50mL CH₂Cl₂. The resulting light brown solution was stirred at rt untilurea formation was complete (˜2 h). To the mixture was then added DBU(4.6 mL, 30 mmol), and the resulting brown colored solution was stirredat rt until hydantoin formation was complete (˜15 h). The product (4.72g, 62%) was collected by filtration and washing with CH₂Cl₂ (2×). Themother liquor was then diluted with CH₂Cl₂ and washed with H₂O (2×), 1 NHCl (2×) and brine. After removal of most of the solvent under reducedpressure, further product (1.2 g, 16%) was collected by filtration andwashing with CH₂Cl₂ (2×). Recrystallization of the 4.72 g of crudeproduct from hot THF and hexane gave 4.5 g of analytically pure product.¹H NMR (DMSO-d₆) δ 2.05-2.11 (m, 1H), 2.15-2.22 (m, 1H), 2.20, 2.24 (s,3H), 3.29-3.33 (m, 1H), 3.59-3.68 (m, 1H), 4.42-4.50 (m, 2H), 5.64, 5.72(d, J=3.9, 3.3, 1H), 7.22, 7.51 (d, J=8.3, 1H), 7.96 (d, J=8.2, 1H); ¹³CNMR (DMSO-d₆) δ 15.4, 15.6, 35.5, 35.6, 43.3, 43.4, 68.8, 69.3, 69.8,112.9, 113.1, 115.8, 128.1, 128.7, 132.1, 136.3, 136.4, 136.9, 137.1,158.6, 169.1, 169.6; HPLC a) column: Phenominex ODS C18 4.6×50 mm, 4 mingradient, 10% MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA; 1 minhold; 4 mL/min, UV detection at 254 nm, 2.07 and 2.32 min retentiontime; HPLC b) column: Shimadzu Shim-Pack VP-ODS C18 (4.6×50 mm), 4 mingradient, 10% MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA, 1 minhold, 4 mL/min, UV detection at 254 nm, 1.93 and 2.23 min retentiontime; Chiral HPLC column: Daicel Chiralcel OD 4.6×250 mm, isocratic, 30min, 25% isopropanol/hexanes, 1 mL/min, UV detection at 254 nm; ShimadzuHPLC: 17.99 min retention time (>99%): Column: Hypercarb 5μ, 4.6×100 mm,25° C., isocratic, 30 min ACN/H₂O (35:65); 1 mL/min, 10.99 min retentiontime; MS (ES) m/z 306 [M+H]⁺. Alternatively, compound 23G can also beprepared by the following procedure: A solution of 22C (0.10 g, 0.28mmol) and copper cyanide (0.03 g, 0.34 mmol) in DMF (1 mL) was refluxedfor 3 h, cooled to rt, and diluted with water. The resulting solid wasfiltered, washed with water, dried and purified using preparative HPLCto afford the title compound (27 mg).

Alternatively, compound 23G can also be prepared by the followingprocedures: A solution of 22C (0.10 g, 0.278 mmol) and copper cyanide(0.03 g, 0.334 mmol) in DMF (1 mL) was refluxed for 3 h, cooled to rtand diluted with water. The resulting solid was filtered, washed withwater, dried and purified using preparative HPLC to afford the titlecompound (27 mg). HPLC: 99% at 2.06, 2.34 min (retention time)(Conditions: Phenom. Lura C18 (4.6×50 mm); Eluted with 0% to 100% B, 4min gradient (A=90% H₂O—10% MeOH—0.1% TFA and B=10% H₂O—90% MeOH—0.1%TFA); Flow rate at 4.0 mL/min. UV detection at 220 nm). Chiral HPLC:retention time=11.04 min (99%); Conditions: OD (4.6×250 mm); Eluted with25% isopropanol in hexane for 30 min at 1 mL/min. MS (ES) m/z 306[M+1]⁺.

EXAMPLE 24(7S,7aR)-2-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

The title compound was prepared from compound 1A and compound 23E in amanner similar to that described in Example 19. mp 237-238° C.; HPLC:100% at 2.11 and 2.36 min (retention time) (Conditions: Phenomenex LunaC18 (4.6×50 mm); Eluted with 0% to 100% B, 4 min gradient (A=90% H₂O—10%MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 4mL/min. UV detection at 220 nm). Chiral HPLC: retention time=15.9 min(100%); Conditions: OD (4.6×250 mm); Eluted with 20% isopropanol inhexane for 30 min at 1 mL/min, MS (ES) m/z 306 [M+1]⁺.

EXAMPLE 25(7R,7aS)-2-Chloro-4-(7-hydroxy-7a-methyl-1,3-dioxotetrahydropyrrolo[2-c]imidazol-2-yl)-3-methylbenzonitrile

To a stirring suspension of compound 23G (600 mg, 1.96 mmol) inanhydrous THF (10 mL) at rt was added DMPU (4 mL). The resulting clearsolution was cooled to −78° C., then a 2.0 M solution of LDA in THF(1.96 mL, 3.92 mmol) was added slowly so that the reaction temperaturewas maintained below −72° C. The resulting dark brown solution wasstirred at −78° C. for 30 min, then iodomethane (0.36 mL, 3.92 mmol) wasadded dropwise. After addition, the reaction was stirred at −78° C. for30 min, then at 0° C. for 3 h. The reaction was quenched with 5% aqueousKHSO₄ and extracted with EtOAc (3×). The combined extracts were washedwith brine, dried (Na₂SO₄), filtered and concentrated. The residue waschromatographed (silica gel), eluting with 1% MeOH in EtOAc/CH₂Cl₂ (1:9)to give a mixture of compound 25 and compound 26 (150 mg), which wasfurther purified using preparative HPLC to afford the title compound (20mg). HPLC: 99% at 4.40, 5.20 min (retention time) (Conditions: Zorbax SBC18 (4.6×75 mm); Eluted with 0% to 100% B, 8 min gradient. (A=90%H₂O—10% MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at2.5 mL/min. UV detection at 220 nm). Chiral HPLC: retention time=7.9 min(99%); Conditions: OD (4.6×250 mm); Eluted with 20% isopropanol inhexane for 30 min at 1 mL/min, MS (ES) m/z 320 [M+1]⁺.

Alternatively, the compound of Example 25 can be prepared in thefollowing manner

25A. (2S,3R)-3-Hydroxy-2-methylpyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester 2-methyl ester

To a solution of (2S,3R)—N-tert-butyloxycarbonyl-3-hydroxy-2pyrrolidinecarboxylic acid methyl ester (1D) (1.13 g, 4.61 mmol) in THF(73 mL) at −78° C. was slowly added a 1.8 M solution of LDA (7.70 mL,13.86 mmol). After stirring for 1 h, iodomethane (2.87 mL, 46.10 mmol)was added, and the reaction was stirred for 1 h at −78° C., beforewarming gradually to −20° C. for 3 h, and then was stored at −40° C.overnight. After quenching with saturated aqueous NH₄Cl, water and EtOAcwere added and the layers were separated. The organic layer was washedwith brine, and the aqueous layer was extracted with EtOAc. The combinedorganics were dried (MgSO₄), filtered and concentrated under reducedpressure. The residue was purified using preparative HPLC (Luna C-18,21.2×250 mm, eluting with 50-90% solvent B (A=90% H₂O—10% MeOH and B=10%H₂O—90% MeOH) over 30 min; Flow rate at 10 mL/min. UV detection at 220nm) to provide a mixture of the starting material and its epimer as ayellow oil (490 mg) and the title compound as a yellow oil (362 mg);LC/MS m/z 260 [M+H]⁺.

25B.(7R,7aS)-2-Chloro-4-(7-hydroxy-7a-methyl-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To a solution of (2S,3R)-3-hydroxy-2-methyl-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester (25A) (87 mg, 0.32 mmol) inCH₂Cl₂ (1.3 mL) at 0° C. was added Hunig's base (111 mL, 0.64 mmol).After stirring for 15 min, 2-chloro-4-isocyanato-3-methylbenzonitrile(23A) was added, and after an additional 10 min, the ice bath wasremoved. The reaction was stirred for 2 h and then diluted with water.The layers were separated and the organic layer was washed with brine,dried (MgSO₄), filtered and concentrated under reduced pressure. Theresulting solid was purified using preparative HPLC (Luna C-18, 21.2×100mm, eluting with 40-100% solvent B (A=90% H₂O—10% MeOH and B=10% H₂O—90%MeOH) over 15 min; Flow rate at 20 mL/min; UV detection at 220 nm) toprovide the title compound (67 mg) as a white film; LC/MS m/z 661[2M+23]⁺.

EXAMPLE 26(7R,7aS)-2-Chloro-3-ethyl-4-(7-hydroxy-7a-methyl-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzonitrile

A mixed product (150 mg) from Experiment 25 was purified usingpreparative HPLC to afford the title compound (50 mg). HPLC: 99% at4.75, 5.54 min (retention time) (Conditions: Zorbax SB C18 (4.6×75 mm);Eluted with 0% to 100% B, 8 min gradient. (A=90% H₂O—10% MeOH—0.1% H₃PO₄and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5 mL/min UV detectionat 220 nm). Chiral HPLC: retention time=6.4 min (99%); Conditions: OD(4.6×250 mm); Eluted with 20% isopropanol in hexane for 30 min at 1mL/min, MS (ES) m/z 334 [M+1]⁺.

EXAMPLE 27(7S,7aR)-2-Chloro-4-(7-hydroxy-7a-methyl-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

The title compound was prepared and isolated as a white solid (30 mg)from compound 24 in a manner similar to that described in Example 25.HPLC: 99% at 4.49, 5.55 min (retention time) (Conditions: Zorbax SB C18(4.6×75 mm); Eluted with 0% to 100% B, 8 min gradient. (A=90% H₂O—10%MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5mL/min UV detection at 220 nm). Chiral HPLC: retention time=8.1 min(99%); Conditions: OD (4.6×250 mm); Eluted with 20% isopropanol inhexane for 30 min at 1 mL/min, MS (ES) m/z 320 [M+1]⁺.

EXAMPLE 28(7S,7aR)-2-Chloro-3-ethyl-4-(7-hydroxy-7a-methyl-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzonitrile

The title compound (35 mg) was obtained as a white solid in the reactionoutlined in Example 27 by a procedure similar to that described inExample 26. HPLC: 99% at 4.79, 5.56 min (retention time) (Conditions:Zorbax SB C18 (4.6×75 mm); Eluted with 0% to 100% B, 8 min gradient.(A=90% H₂O—10% MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flowrate at 2.5 mL/min UV detection at 220 nm). Chiral HPLC: retentiontime=7.1 min (99%); Conditions: OD (4.6×250 mm); Eluted with 20%isopropanol in hexane for 30 min at 1 mL/min, MS (ES) m/z 334 [M+1]⁺.

EXAMPLES 29 to 42

Additional compounds of the present invention were prepared byprocedures analogous to those described above. The compounds of Examples29 to 42 have the following structure:

where G, the compound name, retention time, molecular mass, and theprocedure employed, are set forth in Table 1. The chromatographytechniques used to determine the compound retention times of Table 1 areas follows: LC/MS=Phenom. Luna C18, 4.6×50 mm eluting with 10-90%MeOH/H₂O over 4 min containing 0.1% TFA; 4 mL/min, monitoring at 220 nmThe molecular mass of the compounds listed in Table 1, where provided,were determined by MS by the formula m/z.

TABLE 1 Retention Time (Min)/ Proced. Example Compound Molecular of No.G Name Mass Ex. 29

(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzonitrile2.07LCMS/258 [M + H]⁺ 2 30

(7R,7aS)-3-Ethyl-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzonitrile2.50LCMS/286 [M + H]⁺ 2 31

(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-methoxybenzonitrile2.35LCMS/288 [M + H]⁺ 2 32

(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-propoxybenzonitrile2.76LCMS/316 [M + H]⁺ 2 33

(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-propoxybenzonitrile3.10LCMS/316 [M + H]⁺ 2 34

(7R,7aS)-4-(7-Hydroxy-1,3-dioxo-tetrahydro-pyrrolo[1,2-c]imidazol-2-yl)-3,5-dimethyl-benzonitrile1.44LCMS/286 [M + H]⁺ 2 35

(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2,3-dimethylbenzonitrile1.79, 1.96LCMS/286 [M + H]⁺ 2 36

(7R,7aS)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methoxy-2-methylbenzonitrile1.83LCMS/302 [M + H]⁺ 21  37

(7R,7aS)-3-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-methylbenzonitrile1.71, 2.28LCMS/306 [M + H]⁺ 2 38

(7R,7aS)-2,3-Difluoro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzonitrile3.59LCMS/294 [M + H]⁺ 2 39

(7R,7aS)-2-Chloro-3-fluoro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzonitrile2.12LCMS/310 [M + H]⁺ 20  40

(7R,7aS)-3-Ethyl-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-(trifluoromethyl)-benzonitrile2.48, 2.79LCMS/354 [M + H]⁺ 18  41

(7R,7aS)-2-Fluoro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile3.70LCMS/290 [M + H]⁺ 2 42

(7R,7aS)-6-Cyano-3-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-2-methylbenzoicacidmethyl ester 1.95LCMS/330 [M + H]⁺ 2

EXAMPLE 43(7S,7aR)-4-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylphthalonitrile

A solution of compound 23C (100 mg, 0.327 mmol), CuCN (15 mg, 0.163mmol) and CuBr (23 mg, 0.163 mmol) in DMF (1 mL) was refluxed for 8 h,cooled to rt and diluted with water. The resulting solid was filtered,washed with water, dried and purified using preparative HPLC to affordthe title compound (8 mg). HPLC: 99% at 1.83 min (retention time)(Conditions: Phenom. Lura C18 (4.6×50 mm); Eluted with 0% to 100% B, 4min gradient (A=90% H₂O—10% MeOH—0.1% TFA and B=10% H₂O—90% MeOH—0.1%TFA); Flow rate at 4.0 mL/min UV detection at 220 nm). Chiral HPLC:retention time=21.40 min (99%); Conditions: OD (4.6×250 mm); Eluted with25% isopropanol in hexane for 30 min at 1 mL/min MS (ES) m/z 297 [M+1]⁺.

EXAMPLE 44(7R,7aS)-3-Chloro-5-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)pyridine-2-carbonitrile

44A. 3-Chloropyridine-N-oxide

Commercially available 3-chloropyridine (11.4 g, 100 mmol) was dissolvedin AcOH (60 mL) and 30% hydrogen peroxide (15 mL) was added. The mixturewas heated to 70° C. for 12 h. The cooled mixture was concentrated underreduced pressure. The residue was diluted with chloroform (50 mL) andsolid potassium carbonate (20 g) was added and the mixture stirred for 1h, after which it was filtered and concentrated to give a yellow-greenoil (10.21 g, 79%), which by NMR contained ˜8% of starting material. Theoil was used directly without further purification.

44B. 3-Chloro-2-cyanopyridine

Compound 44A (2.59 g, 20 mmol), trimethylsilylcyanide (5.95 g, 60 mmol),Et₃N (4.05 g, 40 mmol), and acetonitrile (20 mL) were combined in a3-necked 250-mL round-bottomed flask under nitrogen and refluxed for 6h, at which time HPLC analysis indicated complete consumption ofstarting material. The cooled reaction mixture was concentrated underreduced pressure to give a brown semi-solid which was partitionedbetween 3 N aqueous Na₂CO₃ and CH₂Cl₂. The organic layer was washed withwater, brine, dried (MgSO₄), filtered, and concentrated under reducedpressure. The residue was purified by flash chromatography using 5%Et₂O/CH₂Cl₂ as eluent to give the product 44B as a white crystallinesolid (1.84 g, 67%). ¹H NMR (CDCl₃) 7.37 (dd, J=4.7, 8.4 Hz, 1 H), 7.73(dd, J=1.4, 8.2 Hz, 1 H), 8.47 (dd, J=1.3, 4.6 Hz, 1 H); LC/MS m/z 139[M+H]⁺.

44C. 3-Chloro-2-cyano-5-nitropyridine

To a solution of compound 44B (1.75 g, 12.7 mmol) in CH₂Cl₂ (40 mL)cooled at 0-5° C. was added dropwise a CH₂Cl₂ solution (25 mL)containing tetrabutylammonium nitrate (5.02 g, 16.5 mmol) andtrifluoroacetic anhydride (3.15 g, 15 mmol) over 20 min. The reactionwas stirred for 2 h at 0-5° C., warmed to rt and stirred for two days.The mixture was stirred with saturated aqueous Na₂CO₃ for 1 h and theorganic layer was washed with brine and dried (MgSO₄), filtered, andconcentrated under reduced pressure. The residue was purified by flashchromatography using CH₂Cl₂ as eluent to give the product as a yellowsolid (0.43 g, 18%). ¹H NMR (CDCl₃) 8.62 (d, J=2.4 Hz, 1H), 9.33 (d,J=2.3 Hz, 1 H); LC/MS m/z 183 [M+H]⁺.

44D. 5-Amino-3-chloro-2-cyanopyridine

To a solution of Compound 44° C. (0.35 g, 1.9 mmol) in 90% EtOH (10 mL)was added calcium chloride (0.06 g, 0.55 mmol), followed by iron powder(0.56 g, 10 mmol). The resulting mixture was stirred at rt overnight,then filtered through a pad of Celite®, the pad washed with EtOAc, andthe filtrate concentrated under reduced pressure. The residue waspurified by flash chromatography using 10% Et₂O/CH₂Cl₂ to give alight-brown solid (0.13 g, 43%). ¹H NMR (CDCl₃) 7.02 (br s, 2H), 7.28(d, J=2.2 Hz, 1 H), 8.16 (d, J=2.2 Hz, 1 H); LC/MS m/z=154 [M+H]⁺.

44E.(7R,7aS)-3-Chloro-5-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)pyridine-2-carbonitrile

The title compound was synthesized from compound 44D by proceduresanalogous to those described for Examples 2E to 2F. HPLC: 100% at 1.67min. (retention time) (Conditions: Shim-Pak VP-ODS (4.6×50 mm); Elutedwith 0% to 100% B, 4 min gradient, 1 min hold. (A=90% H₂O—10% MeOH—0.1%H₃PO₄ and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 4 mL/min UVdetection at 220 nm); MS(ES) m/z 293 [M+H]⁺.

EXAMPLE 45(7R,7aS)-6-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)quinoline-2-carbonitrile

45A. 6-Nitroquinoline-N-oxide

Commercially available 6-nitroquinoline (1.0 g, 5.6 mmol) was dissolvedin CHCl₃ (30 mL) and mCPBA (1.76 g, 7.8 mmol) was added portionwise andthe reactions stirred at rt for 48 h. The mixture was then washed withsaturated aqueous NaHCO₃, 1 N aqueous NaOH, and 5% aqueous NaHSO₃, dried(Na₂SO₄), filtered and concentrated under reduced pressure to givecompound 45A (1.0 g, 93%) as a light yellow solid. LC/MS m/z 191 [M+H]⁺.45B. 2-Cyano-6-nitroquinoline

To a suspension of compound 45A (400 mg, 2.1 mmol) in MeCN (15 mL) wasadded trimethylsilylcyanide (0.34 mL, 2.5 mmol), followed by a slowaddition of Et₃N (0.65 mL, 4.6 mmol). The reaction mixture was heated to75° C. for 30 min, then cooled to rt, concentrated, dried under vacuum,and purified by silica gel flash chromatography, eluting with CH₂Cl₂ togive the title compound (150 mg, 36%) as a white solid. LC/MS m/z 200[M+H]⁺.

45C. 2-Cyano-6-aminoquinoline

Compound 45B (212 mg, 1.1 mmol) was dissolved in 30 mL of a 1:2 mixtureof EtOAc and MeOH and hydrogenated at 1 atmosphere of hydrogen in thepresence of 10% Pd/C (42 mg, 20 wt. %) for 1 h. The reaction wasfiltered, concentrated and purified by silica gel flash chromatographyeluting with 2-3% MeOH—CH₂Cl₂ to give the title compound (142 mg, 79%)as a light yellow solid. LC/MS m/z 170 [M+H]⁺.

45D.(7R,7aS)-6-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)quinoline-2-carbonitrile

The title compound was prepared from 45C by procedures analogous tothose described in Experiment 2E and 2F. HPLC: 100% at 1.81 min(retention time) (Conditions: YMC S5 C18 (4.6×50 mm); Eluted with 0% to100% B, 8 min gradient, 3 min hold. (A=90% H₂O—10% MeOH—0.1% H₃PO₄ andB=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5 mL/min UV detection at220 nm). LC/MS m/z 309 [M+H]⁺.

EXAMPLE 46(7R,7aS)-7-Hydroxy-2-(2-oxo-1,2-dihydroquinolin-6-yl)-tetrahydropyrrolor[1,2-c]imidazole-1,3-dione

46A. 6-Nitro-2-acetoxyquinoline

A solution of compound 45A (564 mg, 2.96 mmol) in Ac₂O (20 mL) washeated to 145° C. for 5 h. After cooling to rt, the reaction wasconcentrated and the residue purified by silica gel flash chromatographyeluting with CH₂Cl₂ to give the title compound (227 mg, 33%) as a beigesolid. LC/MS m/z 233 [M+H]⁺.

46B. 6-Nitro-2-quinolone

Compound 46A (210 mg, 0.90 mmol) was dissolved in MeOH (10 mL), K₂CO₃was added, and the reaction was stirred at rt for 30 min. The reactionwas then concentrated under reduced pressure and subsequently purifiedby silica gel flash chromatography eluting with 5-10% MeOH/CH₂Cl₂(gradient) to give the title compound (162 mg, 94%) as a pink solid.LC/MS m/z 191 [M+H]⁺.

46C. 6-Nitro-N-benzyl-2-quinolone

Compound 46B (50 mg, 0.26 mmol) was dissolved in DMF (1 mL), CsF (120mg, 0.79 mmol) and benzyl chloride (0.09 mL, 0.79 mmol) were added, andthe reaction was stirred at rt for 16 h. The reaction mixture was thenconcentrated under reduced pressure and subsequently purified by silicagel flash chromatography eluting with 0-5% EtOAc/CH₂Cl₂ (gradient) togive the title compound (57 mg, 77%). LC/MS m/z 281 [M+H]⁺.

46D. 6-Amino-N-benzyl-2-quinolone

The title compound (34 mg) was prepared from compound 46C in a mannersimilar to that described in Experiment 45C. LC/MS m/z 251 [M+H]⁺.

46E.(7R,7aS)-7-Hydroxy-2-(2-oxo-1,2-dihydro-N-benzylquinolin-6-yl)tetrahydropyrrolo[1,2-c]imidazole-1,3-dione

The title compound was prepared from compound 46D by proceduresanalogous to those described in Experiment 2E and 2F. LC/MS m/z 390[M+H]⁺.

46F.(7R,7aS)-7-Hydroxy-2-(2-oxo-1,2-dihydroquinolin-6-yl)tetrahydropyrrolo[1,2-c]imidazole-1,3-dione

Compound 46E (80 mg, 0.20 mmol) was dissolved in MeOH (5 mL) andhydrogenated at 1 atmosphere of hydrogen for 24 h in the presence of 20mg Pd(OH)₂. The reaction was then filtered, concentrated under reducedpressure, and subsequently purified by silica gel flash chromatographyeluting with a gradient of 5%-10% MeOH in CH₂Cl₂ to give the titlecompound (16 mg) as a white solid. HPLC: 98% at 0.79 min (retentiontime) (Conditions: YMC S5 C18 (4.6×50 mm); Eluted with 0% to 100% B, 8min gradient, 3 min hold. (A=90% H₂O—10% MeCN—0.1% TFA and B=10% H₂O—90%MeCN—0.1% TFA); Flow rate at 2.5 mL/min UV detection at 220 nm). LC/MSm/z 300 [M+H]⁺.

EXAMPLE 47(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)biphenyl-2-carbonitrile

47A. 4-Cyano-3-phenylaniline

Commercially available 3-chloro-4-cyanoaniline (305 mg, 2.0 mmol) wasdissolved in NMP (8 mL), and CsF (608 mg, 4.0 mmol), phenyl boronic acid(268 mg, 2.2 mmol) and dichlorobis(tricyclohexylphosphino)palladium(73mg, 0.1 mmol) were added. The reaction was then heated to 100° C. for16 h. After cooling to rt, the reaction mixture was taken up in EtOAc,washed with water (2×), dried (Na₂SO₄), concentrated under reducedpressue, and purified by silica gel flash chromatography eluting withEtOAc/hexanes (1:1) to give compound 47A (371 mg, 96%) as a yellowsolid. LC/MS m/z 195 [M+H]⁺.

47B.(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)biphenyl-2-carbonitrile

The title compound was prepared from compound 47A by proceduresanalogous to those described in Experiment 2E and 2F. HPLC: 96% at 3.24min (retention time) (Conditions: YMC S5 C18 (4.6×50 mm); Eluted with 0%to 100% B, 8 min gradient, 3 min hold. (A=90% H₂O—10% MeOH—0.1% H₃PO₄and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5 mL/min UV detectionat 220 nm). LC/MS m/z 334 [M+H]⁺.

EXAMPLE 48(7R,7aS)-4′-Fluoro-5-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)biphenyl-2-carbonitrile

48A. 4-Cyano-3-(4-fluorophenyl)aniline

The title compound was prepared from commercially available3-chloro-4-cyanoaniline in a similar fashion to that described inExperiment 47A and isolated as a yellow solid. LC/MS m/z 213 [M+H]⁺.

48B.(7R,7aS)-4′-Fluoro-5-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)biphenyl-2-carbonitrile

The title compound was prepared from compound 48A by proceduresanalogous to those described in Experiment 2E and 2F. HPLC: 99% at 3.32min (retention time) (Conditions: YMC S5 C18 (4.6×50 mm); Eluted with 0%to 100% B, 8 min gradient, 3 min hold. (A=90% H₂O—10% MeOH—0.1% H₃PO₄and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5 mL/min. UVdetection at 220 nm.). LC/MS m/z 352 [M+H]⁺.

EXAMPLE 49(7R,7aS)-5-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-4′-methoxybiphenyl-2-carbonitrile

49A. 4-Cyano-3-(4-methoxyphenyl)aniline

The title compound was prepared from commercially available3-chloro-4-cyanoaniline in a manner similar to that described inExperiment 47A and isolated as a yellow solid. LC/MS m/z 225 [M+H]⁺.

49B.(7R,7aS)-5-(7-Hydroxy-1,3-dioxo-tetrahydro-pyrrolo[1,2-c]imidazol-2-yl)-4′-methoxybiphenyl-2-carbonitrile

The title compound was prepared from compound 49A by proceduresanalogous to those described in Experiment 2E and 2F. HPLC: 96% at 3.24min (retention time) (Conditions: YMC S5 C18 (4.6×50 mm); Eluted with 0%to 100% B, 8 min gradient, 3 min hold. (A=90% H₂O—10% MeOH—0.1% H₃PO₄and B=10% H₂O—90% MeOH—0.1% H₃PO₄); Flow rate at 2.5 mL/min UV detectionat 220 nm). LC/MS m/z 364 [M+H]⁺.

EXAMPLE 50(7R,7aR)-7-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzo[1,2,5]thiadiazole-4-carbonitrile

50A. 4-Cyano-7-amino-benzothiadiazole

A solution of 2-cyano-5-nitrophenylenediamine (78 mg, 0.44 mmol,prepared as described in WO 0076501) in SOCl₂ (2 mL) was heated toreflux for 3 h. The resulting mixture was allowed to cool to rt and wasthen poured into ice/water. CH₂Cl₂ was added, the layers were separatedand the aqueous layer was extracted twice with CH₂Cl₂. The combinedorganic phases were dried (MgSO₄), concentrated under reduced pressureand purified by flash chromatography on silica gel eluting with 50%EtOAc in hexanes to give 4-cyano-7-nitrobenzothiadiazole. This materialwas dissolved in AcOH (2 mL) containing EtOAc (1 mL) and H₂O (0.2 mL)and heated to 70° C. At this temperature, iron powder (78 mg, 1.41 mmol)was added in one portion and the dark mixture was stirred for 20 minbefore cooling to rt. The reaction mixture was then filtered through apad of Celite® eluting with EtOAc, washed with saturated Na₂CO₃solution, dried (MgSO₄) and concentrated under reduced pressure.Purification by flash chromatography on silica gel eluting with 20-70%EtOAc in hexanes afforded the title compound (47 mg, 67%) as a brownsolid.

50B.(7R,7aR)-7-(7-Hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzo[1,2,5]thiadiazole-4-carbonitrile

The title compound (12.2 mg) was prepared from compound 50A byprocedures analogous to those described in Experiment 2E and 2F. HPLC:99% at 1.57 min (retention time) (Conditions: Phenom. Luna. (4.6×50 mm);Eluted with 0% to 100% B, 4 min gradient, 1 min hold. (A=90% H₂O—10%MeOH—0.1% TFA and B=10% H₂O—90% MeOH—0.1% TFA); Flow rate at 4.0 mL/minUV detection at 220 nm). Chiral HPLC: retention time=22.68 min (98%);Conditions: OD (4.6×250 mm); Eluted with 25% isopropanol in hexane for30 min at 1 mL/min; MS (ES) m/z 316 [M+1]⁺.

EXAMPLE 51(7R,7aR)-2-Chloro-4-(7-hydroxy-3-oxotetrahydropyrrolo[1,2-c]imidazol-2-yl)3-methylbenzonitrile

To a solution of compound 23G (150 mg, 0.5 mmol) in anhydrous THF (12mL) cooled to −78° C. was added dropwise 1.0 M LiEt₃BH solution in THF(0.5 mL, 0.5 mmol). After addition, the reaction was stirred at −78° C.for 4 h, then quenched by addition of saturated aqueous Na₂CO₃ (5 mL).The reaction was warmed to 0° C., 30% H₂O₂ (˜0.5 mL) was added, and thereaction was stirred at 0° C. for 30 min, then extracted with CH₂Cl₂(3×). The combined extracts were washed with brine, dried (Na₂SO₄),filtered, and concentrated under reduced pressure overnight. The residue(˜120 mg) was dissolved in anhydrous CH₂Cl₂, and to the resultingsolution cooled to −78° C. was added dropwise triethylsilane (0.5 mL,3.1 mmol), followed by boron trifluoride diethyl etherate (0.5 mL, 3.9mmol). The reaction mixture was stirred at −78° C. for 2 h, thenadditional triethylsilane (0.3 mL, 1.88 mmol) and boron trifluoridediethyl etherate (0.3 mL, 2.35 mmol) were added, and the reaction wasstirred at 0° C. overnight. The reaction was then quenched withsaturated aqueous Na₂CO₃ (10 mL), then extracted with CH₂Cl₂ (3×). Thecombined extracts were washed with brine, dried (Na₂SO₄), filtered, andconcentrated under reduced pressure to give a crude product (˜100 mg).The crude product was purified using preparative HPLC to give 30 mg,which was further purified using chiral preparative HPLC to afford thetitle compound (9 mg). HPLC: 98% at 4.39 min (retention time)(Conditions: Zorbax SB C18 (4.6×75 mm); Eluted with 0% to 100% B, 8 mingradient (A=90% H₂O—10% MeOH—0.1% H₃PO₄ and B=10% H₂O—90% MeOH—0.1%H₃PO₄); Flow rate at 2.5 mL/min UV detection at 220 nm). Chiral HPLC:retention time=10.59 min (99%); Conditions: OD (4.6×250 mm); Eluted with20% isopropanol in hexane for 30 min at 1 mL/min; MS (ES) m/z 292[M+1]⁺.

Alternatively, the Compound of Example 51 can be prepared by thefollowing sequence:

51A.(2S,3R)-3-(tert-Butyldimethylsilanyloxy)pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester

To a solution of (2S,3R)—N-tert-butyloxycarbonyl-3-hydroxy-2pyrrolidinecarboxylic acid methyl ester (1D) (0.63 g, 2.56 mmol) inCH₂Cl₂ (12 mL) at rt was added imidazole (0.35 g, 5.14 mmol), and thentert-butydimethylsilyl chloride (0.43 g, 2.83 mmol). After stirring for3 h, the reaction mixture was partitioned between H₂O and CH₂Cl₂. TheCH₂Cl₂ layer was washed with 1 M H₃PO₄, NaHCO₃ and brine, dried (MgSO₄),then filtered and concentrated under reduced pressure. The residue waschromatographed (silica gel) eluting with 30% EtOAc/hexane to yield thetitle compound (0.91 g). LC/MS m/z 360 [M+H]⁺.

51B.(2R,3R)-3-(tert-Butyldimethylsilanyloxy)-2-hydroxymethylpyrrolidine-1-carboxylicacid tert-butyl ester

To(2S,3R)-3-(tert-Butyl-dimethyl-silanyloxy)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester (51A) (8.05 g, 22.39 mmol) in THF(90 mL) at −78° C. was added a 1 M solution of Super-Hydride® in THF(112 mL, 112 mmol) in five portions over 15 min The cold bath wasremoved and the reaction was allowed to warm to rt. After 3 h, thereaction was poured into a 1-L Erlenmeyer flask and was carefullyquenched with ice while stirring and then diluted with EtOAc. The layerswere separated and the organic layer washed with 1 M H₃PO₄, NaHCO₃ andbrine, dried (MgSO₄), filtered and concentrated. The residue was dilutedwith CH₂Cl₂, stirred with silica gel overnight, then concentrated andpurified via flash chromatography eluting with 30% EtOAc/hexane toobtain the title compound (6.70 g) as a clear oil.

51C.(2S,3R)-3-(tert-Butyldimethylsilanyloxy)-2-formylpyrrolidine-1-carboxylicacid tert-butyl ester

To(2R,3R)-3-(tert-Butyl-dimethyl-silanyloxy)-2-hydroxymethyl-pyrrolidine-1-carboxylicacid tert-butyl ester (51B) (6.70 g, 20.24 mmol) in CH₂Cl₂ (100 mL) at0° C. was added Dess-Martin periodinane. The ice bath was removed andthe reaction was warmed to rt. After 2 h, saturated aqueous Na₂S₂O₃ andNaHCO₃ (ca. 100 mL each) were added and the reaction mixture was stirredvigorously for 0.5 h. The layers were separated, the organic layer waswashed with a mixture of saturated aqueous Na₂S₂O₃ and NaHCO₃ followedby brine, dried (MgSO₄), and was then filtered and concentrated toobtain the title compound (7.20 g) as a yellow oil.

51D.(2R,3R)—N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsiloxy)-2-[(3-chloro-4-cyano-2-methylphenylamino)methyl]pyrrolidine

To a solution of(2S,3R)-3-(tert-Butyldimethylsilanyloxy)-2-formyl-pyrrolidine-1-carboxylicacid tert-butyl ester (51C) (661 mg, 2.0 mmol) in 5% DMF in CH₂Cl₂ (10mL) at rt was added 4-amino-2-chloro-3-methyl-benzonitrile (340 mg, 2.04mmol) followed by NaBH(OAc)₃ (636 mg, 3.0 mmol) and HOAc (180 μL, 3mmol). The reaction was stirred under nitrogen at rt for 18 h.Additional portions of NaBH(OAc)₃ (424 mg, 2.0 mmol) and HOAc (120 μL, 2mmol) were added, and the reaction was stirred for an additional 18 h.The reaction was diluted with EtOAc (50 mL) and the organic layer waswashed with saturated aqueous NaHCO₃ (50 ml), dried (MgSO₄), filteredand concentrated. Purification via flash chromatography (silica gel, 0to 15% EtOAc/hexanes) provided the title compound (605 mg, 1.26 mmol,63%). MS m/z 480 [M+H]⁺.

51E.(2R,3R)-3-(tert-Butyldimethylsilanyloxy)-2-[(3-chloro-4-cyano-2-methylphenylamino)methyl]pyrrolidine-1-carboxylicacid tert-butyl ester

(2R,3R)—N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsiloxy)-2-[(3-chloro-4-cyano-2-methylphenylamino)methyl]pyrrolidine(51D) (35 mg, 0.07 mmol) was dissolved in 50% TFA/CH₂Cl₂ and stirred for2 h then concentrated. The residue was dissolved in EtOAc, saturatedaqueous NaHCO₃ was added and the reaction mixture was stirred vigorouslyfor 0.5 h. The layers were separated, and the organic layer was washedwith NaHCO₃ and brine, dried (MgSO₄), filtered and concentrated toprovide the title compound (22 mg) as a yellow film. LC/MS m/z 380[M+H]⁺.

51F.(7R,7aR)-2-Chloro-4-(7-hydroxy-3-oxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To a solution of(2R,3R)-3-(tert-Butyldimethylsilanyloxy)-2-[(3-chloro-4-cyano-2-methylphenylamino)methyl]pyrrolidine-1-carboxylicacid tert-butyl ester (51E) (22 mg, 0.06 mmol) dissolved in THF (1 mL)was added 1,1′-carbonyldiimidazole (9.40 mg, 0.06 mmol) and the mixturewas stirred at rt for 3 days, and then brought to reflux for 1 h. Analiquot (˜half of the reaction mixture) was treated with DBU (10 μL,0.07 mmol) and the reaction mixture was heated at reflux overnight. Thereaction was then diluted with EtOAc and water, and the layers wereseparated. The organic layer was washed with brine, concentrated underreduced pressure, and purified via preparative HPLC (Luna C-18, 21.2×100mm, eluting with 60-100% solvent B (A=90% H₂O—10% MeOH—0.1% TFA andB=10% H₂O—90% MeOH—0.1% TFA) over 12 min; Flow rate at 20 mL/min. UVdetection at 220 nm). The major peak was collected, concentrated andtreated with TFA (2 mL) overnight. The reaction was concentrated andpurified via preparative HPLC (Luna C-18, 21.2×100 mm, eluting with40-100% solvent B (A=90% H₂O—10% MeOH—0.1% TFA and B=10% H₂O—90%MeOH—0.1% TFA) over 10 min; Flow rate at 20 mL/min. UV detection at 220nm) to provide the title compound (1 mg). LC/MS m/z 292 [M+H]⁺.

EXAMPLE 522-Chloro-4-[3,7-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To a solution of compound 51 (0.02 g, 0.07 mmol) in CH₂Cl₂ (0.5 mL) wasadded a solution of Dess-Martin periodinane (0.038 g, 0.086 mmol) at rt.The reaction mixture was stirred at rt for 2 h. The reaction mixture waswashed with NaHCO₃ (2×2 mL), brine, and dried (Na₂SO₄). The CH₂Cl₂ layerwas then evaporated and the crude product was purified by preparativeHPLC to yield 0.01 g of the title compound as a white powder. HPLC: 99%at 1.92 min; Conditions: Phenom. Luna C18 (4.6×50 mm); eluted with 0% to100% B; 4 min gradient (A=90% H₂O—10% ACN—0.1% H₃PO₄ and B=10% H₂O—90%ACN—0.1% H₃PO₄; flow rate at 4 mL/min., UV detection at 220 nm. ChiralHPLC: 98% at 34.2 min; Conditions: (CHIRALPAK® OD column 4.6×250 mm; 25%isopropanol in hexane over 40 min at flow rate 1.0 mL/min, UV detectionat 220 nm); MS (ES) m/z 299 [M+1]⁺.

EXAMPLES 53a and 53b2-Chloro-4-(8-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyridin-2-yl)-3-methylbenzonitrile

53A. (±)-cis-3-hydroxypyridine-2-carboxylic acid

A sample of Rh(OH)₃ was prepared according to the procedure described inTetrahedron Lett. 1967, 17, 1663-1664. The3-hydroxypyridine-2-carboxylic acid (0.5 g, 3.6 mmol) was dissolved inaqueous NH₄OH and then added H₂O in a ratio of 1 to 7. Rh(OH)₃ (0.2 g)was added and the reaction mixture was stirred at rt under 70-80 psi ofH₂ for 4 h. The catalyst was filtered through a cake of celite and thefiltrate was evaporated under reduced pressure to afford compound 53A(0.50 g) as a white foam.

53B. (±)-cis-3-Hydoxypiperidine-2-carboxylic acid methyl ester

Hydrogen chloride gas was bubbled through a suspension of3-hydroxy-piperidine-2-carboxylic acid (0.54 g, 0.370 mol) in MeOH (100mL) cooled to 0° C. for 10 min. The resulting clear solution was stirredat rt for 4 h, then evaporated carefully under reduced pressure (whiteprecipitates formed during the concentration). The resulting white solidwas dried overnight under vacuum to afford 0.86 g of the title compoundas an off-white powder.

53C.2-Chloro-4-(8-hydroxy-1,3-dioxohexahydroimidazo[1,5-a]pyridin-2-yl)-3-methylbenzonitrile

To a suspension of compound 53B (0.20 g, 0.77 mmol) in CH₂Cl₂ (3 mL)cooled to 0° C. was added i-Pr₂EtN (0.178 mL, 1.00 mmol). After stirringat 0° C. for 20 min, compound 23E (0.095 g, 0.59 mmol) in CH₂Cl₂ (1 mL)solution was added, along with 4 Å molecular sieves (0.5 g), and theresulting mixture stirred at rt until urea formation was completed (˜2h). The mixture was then stirred at rt until hydantoin formation wascomplete (˜15 h). The reaction mixture was loaded on a silica gelcolumn, eluted with 40% EtOAc/hexane, and 5% MeOH in EtOAc/hexane (1:1)to afford 0.11 g of the title compound as an off-white powder. HPLC: 99%at 1.67-1.79 min; Conditions: Phenom. Luna C18 (4.6×50 mm); Eluted with0% to 100% B; 4 min gradient (A=90% H₂O—10% ACN—0.1% H₃PO₄ and B=10%H₂O—90% ACN—0.1% H₃PO₄), Flow rate at 4 mL/min., UV detection at 220nm). The compound was further loaded on a Chiral AD column, eluted with25% isopropanol in hexane isocratic to afford 50 mg each of enantiomer53a (isomer A; retention time=14.2 min; 100% e.e.) and enantiomer 53b(isomer B; retention time=20 min; 100% e.e.) of the title compound aswhite powders. Chiral HPLC Conditions: (CHIRALPAK® AD column 4.6×250 mm;25% isopropanol in hexane over 30 min at flow rate 1.0 mL/min, UVdetection at 220 nm); MS (ES) m/z 320 [M+1]⁺.

EXAMPLE 54(7S,7aS)-2-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

54A.(2S,3S)-1-(3-Chloro-4-cyano-2-methylphenylcarbamoyl)-3-hydroxy-pyrrolidine-2-carboxylicacid methyl ester.

To a suspension of trans-3-hydroxyproline methyl ester, HCl salt (207mg, 1.14 mmol) in 1.5 mL of CH₂Cl₂ cooled to 0° C. was addeddiisopropylethylamine (0.23 mL, 1.30 mmol) followed by a suspension ofisocyanate 23E (200 mg, 1.04 mmol) in 2 mL of CH₂Cl₂. The suspension wasallowed to warm to rt and stir for 1 h. The reaction mixture was thenwashed with water, and a solid precipitated which was filtered and driedunder vacuum to afford the title compound (210 mg) as a white solid. ¹HNMR (CD₃OD) 1.98-2.01 (m, 1H), 2.13-2.17 (m, 1H), 2.30 (s, 3H),3.69-3.72 (m, 5H), 4.36 (br s, 1H), 4.40 (br s, 1H), 7.41 (d, J=8.80,1H), 7.55 (d, J=8.25, 1H); ¹³C NMR (CD₃OD) 15.88, 33.72, 45.64, 52.94,69.50, 74.55, 110.62, 117.41, 125.65, 132.36, 134.44, 137.83, 144.33,156.43, 172.88; HPLC a) column: Phenominex C18 4.6×50 mm, 4 mingradient, 10% MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA; 1 minhold, 4 mL/min UV detection at 220 nm, 2.30 min retention time; HPLC b)column: Shimadzu Shim-Pack VP-ODS C18 4.6×50 mm, 4 min gradient, 10%MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA, 1 min hold; 4mL/min, UV detection at 220 nm, 2.13 min retention time (97%); HPLC c)column: Daicel Chiralcel OD 4.6×250 mm, Isocratic 25%Isopropanol/Hexanes, 30 min, 1 mL/min, UV detection at 220 nm, 9.03 minretention time (98%); MS (ES) m/z 338 [M+H]⁺.

54B.(2S,3S)-1-(3-Chloro-4-cyano-2-methylphenylcarbamoyl)-3-hydroxy-pyrrolidine-2-carboxylicacid

A suspension of ester 54A (260 mg, 0.770 mmol) in 20 mL of 1.6 N NaOHwas stirred at rt for 45 min. The reaction mixture was acidified to pH 2with 10% HCl and extracted with EtOAc (3×). The combined organic layerswere dried (MgSO₄), filtered and the filtrate concentrated under reducedpressure to afford the title compound (260 mg) as a beige solid. Aportion (50 mg) of the residue was purified by preparative HPLC (reversephase silica gel, 10% MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1%TFA) to afford the title compound (25 mg) as a colorless oil. ¹H NMR(CD₃OD) 2.02-2.05 (m, 1H), 2.17-2.22 (m, 1H), 2.35 (s, 3H), 3.73-3.76(m, 2H), 4.40 (br s, 1H), 4.49 (br s, 1H), 7.47 (d, J=8.80, 1H), 7.58(d, J=8.25, 1H); ¹³C NMR (CD₃OD) 15.83, 33.59, 45.61, 69.53, 74.73,110.40, 117.45, 125.45, 132.34, 137.80, 144.42, 156.50, 159.95, 173.94;HPLC a) column: Phenominex ODS C18 4.6×50 mm, 4 min gradient, 10%MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA; 1 min hold, 4 mL/minUV detection at 220 nm, 1.97 min retention time; HPLC b) column:Shimadzu Shim-Pack VP-ODS C18 4.6×50 mm, 4 min gradient, 10% MeOH/90%H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA, 1 min hold; 4 mL/min, UVdetection at 220 nm, 1.79 min retention time (92%); HPLC c) column:Daicel Chiralcel OD 4.6×250 mm, Isocratic 25% isopropanol/hexanes, 30min, 1 mL/min, UV detection at 220 nm, 6.96 min retention time (98%); MS(ES) m/z 324 [M+H]⁺.

54C.(7S,7aS)-2-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile.

To a suspension of acid 54B (210 mg, 0.649 mmol) in 15 mL ofacetonitrile at rt was added DCC (134 mg, 0.649 mmol) followed byp-nitrophenol (180 mg, 1.30 mmol). The suspension was refluxed for 1 h,cooled to rt and filtered. The filtrate was concentrated under reducedpressure and the residue dissolved in EtOAc, washed with water andbrine, dried (MgSO₄), filtered and concentrated under reduced pressure.The residue was purified by flash chromatography (silica gel,EtOAc/hexanes, 75:25) to afford the title compound (121 mg) as a whitefoam. ¹H NMR (CD₃OD) 1.94-2.05 (m, 1H), 2.18, 2.22 (s, 3H), 2.24-2.33(m, 1H), 3.34-3.40 (m, 1H), 3.67-3.75 (m, 1H), 4.08, 4.16 (d, J=6.05,1H), 4.35, 4.42 (m, 1H), 7.33, 7.38 (d, J=8.25, 1H), 7.70 (m, 1H); ¹³CNMR (CD₃OD) 15.90, 16.11, 36.95, 37.05, 44.88, 45.06, 70.91, 70.94,72.46, 72.90, 115.19, 115.53, 116.60, 116.66, 129.30, 129.46, 132.74,133.03, 137.41, 137.61, 138.08, 138.37, 138.42, 138.53, 159.69, 159.97,172.10, 172.43; HPLC a) column: Phenominex LUNA C18 4.6×50 mm, 4 mingradient, 10% MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA; 1 minhold, 4 mL/min UV detection at 220 nm, 2.32 min retention time; HPLC b)column: Shimadzu Shim-Pack VP-ODS C18 4.6×50 mm, 4 min gradient, 10%MeOH/90% H₂O/0.1% TFA to 90% MeOH/10% H₂O/0.1% TFA, 1 min hold; 4mL/min, UV detection at 220 nm, 2.15 min retention time (100%); HPLC c)column: Daicel Chiralcel OD 4.6×250 mm, Isocratic 25%Isopropanol/Hexanes, 30 min, 1 mL/min, UV detection at 220 nm, 17.65 minretention time (99%); MS (ES) m/z 306 [M+H]⁺.

EXAMPLE 552-Chloro-4-[(7S,7aR)-7-hydroxy-3-oxo-tetrahydro-1H-pyrrolo[1,2-c]imidazol-2(3H)-yl]-3-methylbenzonitrile

55A.Methyl-(2S,3S)-1-{[(3-Chloro-4-cyano-2-methylphenyl)amino]-carbonyl}-3-hydroxypyrrolidine-2-carboxylate

A solution of ester 1A (500 mg, 2.75 mmol) in dry CH₂Cl₂ (10 mL) wascooled to 0° C., treated with Hunig's base (0.53 mL, 3.04 mmol) andstirred at 0° C. for 30 min. The solution was treated with isocyanate23E (505 mg, 2.62 mmol), and the resulting suspension was stirred at rtfor 3 h. The insoluble solids were filtered off and the filtrate waspartitioned between aqueous NH₄Cl (6.0 mL) and CH₂Cl₂ (3×60 mL). Thecombined organic phases were washed with brine, dried (Na₂SO₄), filteredand concentrated under reduced pressure. The residue and insolublesolids were combined and chromatographed (silica gel; EtOAc/hexanegradient) to yield the title compound (177.2 mg, 75.3%) as a whitesolid, mp 189-191° C. HPLC: 1.72 min (retention time) (Conditions: YMCS-5 C-18 (4.6×50 mm), eluting with 0-100% B, 4 min gradient. (A=90%H₂O—10% CH₃CN—0.1% TFA and B=10% H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4mL/min. UV detection at 220 nm). Chiral HPLC: retention time=22.2 min(100%); Conditions: AD (4.6×250 mm); Eluted with 20% isopropanol inheptane for 30 min at 1 mL/min. MS (ES) m/z 338 [M+H]⁺.

55B.Methyl-(2S,3S)-1-{[(3-chloro-4-cyano-2-methylphenyl)amino]-carbonyl}-3-(tert-butyl-dimethylsilanyloxy)-pyrrolidine-2-carboxylate

A cooled (0° C.) solution of compound 55A (510.1 mg, 1.51 mmol) andimidazole (516 mg, 7.58 mmol) in dry DMF (2.6 mL) was treated with 97%tert-butyldimethylsilyl chloride (572 mg, 3.68 mmol), stirred at 0° C.for 5 min then at rt for 24 h. Methanol (3.5 mL) was added and thesolution stirred at rt for another 24 h. The mixture was partitionedbetween 10% citric acid (5.3 mL) and EtOAc (3×50 mL). The combinedorganic phases were washed with brine, dried (Na₂SO₄), filtered andconcentrated under reduced pressure. The resulting syrup waschromatographed (silica gel; EtOAc/hexane gradient) to yield the titlecompound (732.3 mg, 100%) as a white solid, mp 123-125° C. HPLC: 3.38min (retention time) (Conditions: YMC S-5 C-18 (4.6×50 mm), eluting with0-100% B, 4 min gradient. (A=90% H₂O—10% CH₃CN—0.1% TFA and B=10%H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4 mL/min. UV detection at 220 nm.Chiral HPLC: retention time=9.30 min (98.6%); Conditions: AD (4.6×250mm); Eluted with 20% isopropanol in heptane for 30 min at 1 mL/min. MS(ES) m/z 452 [M+H]⁺.

55C.(2R,3S)—N-(3-Chloro-4-cyano-2-methylphenyl)3-(tert-butyldimethylsilanyloxy)-2-(hydroxymethyl)pyrrolidine-1-carboxamide

To a solution of compound 55B (300 mg, 0.66 mmol) in anhydrous THF (6.7mL) at −25° C. was added 1 N LAH in THF (1.34 mL, 1.34 mmol) over aperiod of 10 min. The solution was then warmed to 0° C. and stirred for2.0 h before quenching with H₂O (0.05 mL), 15% NaOH (0.05 mL) and H₂O(0.16 mL). After warming to rt, the mixture was extracted with EtOAc(2×40 mL). The combined organic phases were washed with brine, dried(Na₂SO₄), filtered and concentrated under reduced pressure. Theresultant off-white solid was chromatographed (silica gel; EtOAc/hexanegradient) to yield the title compound (217.2 mg, 78%) as a white solid,mp 174-176° C. HPLC: 3.23 min (retention time) (Conditions: YMC S-5 C-18(4.6×50 mm), eluting with 0-100% B, 4 min gradient. (A=90% H₂O—10%CH₃CN—0.1% TFA and B=10% H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4 mL/min.UV detection at 220 nm. Chiral HPLC: retention time=7.11 min (96.1%);Conditions: AD (4.6×250 mm); Eluted with 20% isopropanol in heptane for30 min at 1 mL/min. MS (ES) m/z 424 [M+H]⁺.

55D.2-Chloro-4-[(7S,7aR)-7-(tert-butyldimethylsilanyloxy)-3-oxo-tetrahydro-1H-pyrrolo[1,2c]imidazol-2-(3H)yl]methylbenzonitrile

A solution of compound 55C (150 mg, 0.35 mmol) in anhydrous THF (4.8 mL)at 0° C. was treated with 97% tert-BuOK (104.2 mg, 0.86 mmol) andstirred at 0° C. for 5 min. To the solution was added a solution oftoluenesulfonyl chloride (81.6 mg, 0.43 mmol) in anhydrous THF, and themixture was stirred at 0° C. for another 10 min as described in by TaekHeon Kim and Gue-Jae Lee J. Org. Chem. 64, 2941-2943 (1999). Thereaction mixture was then quenched with H₂O (4.8 mL), removed from thebath and extracted with EtOAc (2×15 mL). The combined organic phaseswere dried (Na₂SO₄), filtered and concentrated under reduced pressure.The residue was chromatographed (silica gel; EtOAc/hexane gradient) toyield the title compound (128.9 mg, 90%) as a white foam. HPLC: 3.61 min(retention time) (Conditions: YMC S-5 C-18 (4.6×50 mm), eluting with0-100% B, 4 min gradient. (A=90% H₂O—10% CH₃CN—0.1% TFA and B=10%H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4 mL/min. UV detection at 220 nm.Chiral HPLC: retention time=9.54 min (99.3%); Conditions: AD (4.6×250mm); Eluted with 20% isopropanol in heptane for 30 min at 1 mL/min. MS(ES) m/z 406 [M+H]⁺.

55E.2-Chloro-4-[(7S,7aR)-7-hydroxy-3-oxotetrahydro-1H-pyrrolo[1,2-c]imidazol-2(3H)-yl]-3-methylbenzonitrile.

To a solution of compound 55D (112.4 mg, 0.28 mmol) in anhydrous THF(5.0 mL) at 0° C. was added 1.0 M TBAF in THF (0.32 mL, 0.32 mmol). Thesolution was stirred at 0° C. for 10 min, at rt for 19 h, thenpartitioned between 25% NH₄Cl (7.0 mL) and EtOAc (3×40 mL). The combinedorganic phases were washed with brine, dried (Na₂SO₄), filtered andconcentrated under reduced pressure. The resultant off-white solid waschromatographed (silica gel; EtOAc/hexane gradient) to yield the titlecompound (77.4 mg, 95%) as a white solid, mp 148-149° C. HPLC: 5.4 min(retention time) (Conditions: Zorbax C-18 (4.6×75 mm), eluting with0-100% B, 8 min gradient. (A=90% H₂O—10% CH₃OH—0.2% H₃PO₄ and B=10%H₂O—90% CH₃OH—0.2% H₃PO₄); Flow rate at 2.5 mL/min. UV detection at 220nm. Chiral HPLC: retention time=19.9 min (99.7%); Conditions: AD(4.6×250 mm); Eluted with 20% isopropanol in heptane for 30 min at 1mL/min. MS (ES) m/z 290 [M−H]⁻.

EXAMPLE 562-Chloro-4-[(6R,7aS)-6-hydroxy-3-oxotetrahydropyrrolo[1,2-c]imidazol-2(3H)-yl]-3-methylbenzonitrile.

56A. 4-Hydroxypyrrolidine-2-carboxylic acid methyl ester, hydrochloridesalt

The title compound was prepared from trans-4-hydroxy-L-proline (5.0 g,38.1 mmol) by procedures analogous to those described for Example 1A toafford a white solid (6.97 g, 100%), mp 164-166° C.

56B. Methyl(2S,4R)-1-{[(3-Chloro-4-cyano-2-methylphenyl)amino]-carbonyl}-4-hydroxypyrrolidine-2-carboxylate

The title compound was prepared from compound 56A (300 mg, 1.65 mmol)and isocyanate 23E (312 mg, 1.62 mmol) by procedures analogous to thatdescribed for Example 55A to afford a white foam (375.6 mg, 69%). HPLC:1.68 min (retention time) (Conditions: YMC S-5 C-18 (4.6×250 mm),eluting with 0-100% B, 4 min gradient. (A=90% H₂O—10% CH₃CN—0.1% TFA andB=10% H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4 mL/min. UV detection at220 nm. Chiral HPLC: retention time=12.92 min (98.9%); Conditions: AD(4.6×250 mm); Eluted with 20% isopropanol in heptane for 30 min at 1mL/min. MS (ES) m/z 338 [M+H]⁺.

56C. Methyl(2S,4R)-1-{[(3-chloro-4-cyano-2-methylphenyl)amino]-carbonyl}4-(tert-butyldimethylsilanyloxy)pyrrolidine-2-carboxylate.

The title compound was prepared from compound 56B (150 mg, 0.44 mmol) ina manner analogous to that described for compound 55B to afford a whitesolid (156.3 mg, 79%), mp 129-131° C. HPLC: 3.38 min (retention time)(Conditions: YMC S-5 C-18 (4.6×250 mm), eluting with 0-100% B, 4 mingradient. (A=90% H₂O—10% CH₃CN—0.1% TFA and B=10% H₂O—90% CH₃CN—0.1%TFA); Flow rate at 4 mL/min. UV detection at 220 nm. Chiral HPLC:retention time=9.80 min (99.9%); Conditions: AD (4.6×250 mm); Elutedwith 20% isopropanol in heptane for 30 min at 1 mL/min. MS (ES) m/z 452[M+H]⁺.

56D.(2S,4R)—N-(3-Chloro-4-cyano-2-methylphenyl)-4-(tert-butyldimethylsilanyloxy)-2-(hydroxymethyl)pyrrolidine-1-carboxamide

To a solution of compound 56C (145 mg, 0.32 mmol) in anhydrous THF (3.0mL) at 0° C. was added dropwise a solution of 2 M LiBH₄ in THF (0.24 ml,0.48 mmol) as described by Terry Rosen et. al. J. Med. Chem. 31 (8),1598-1611 (1988). The solution was stirred at 0° C. for 2.5 h, quenchedwith 1.0 M K₂CO₃ (1.0 mL) and extracted with EtOAc (2×25 mL). Thecombined organic phases were washed with 1 M K₂CO₃, brine, dried(Na₂SO₄), filtered and concentrated under reduced pressure. The residuewas chromatographed (silica gel; EtOAc/hexane gradient) to yield thetitle compound (125.4 mg, 93%) as a white solid, mp 169-171° C. HPLC:3.31 min (retention time) (Conditions: YMC S-5 C-18 (4.6×250 mm),eluting with 0-100% B, 4 min gradient. (A=90% H₂O—10% CH₃CN—0.1% TFA andB=10% H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4 mL/min. UV detection at220 nm. Chiral HPLC: retention time=7.01 min (99.96%); Conditions: AD(4.6×250 mm); Eluted with 20% isopropanol in heptane for 30 min at 1mL/min. MS (ES) m/z 424 [M+H]⁺.

56E.2-Chloro-4-[(6R,7aS)-6-(tert-butyldimethylsilanyloxy)-3-oxotetrahydro-1H-pyrrolo[1,2-c]imidazol-2(3H)-yl]-3-methylbenzonitrile

The title compound was prepared from compound 56D (124 mg, 0.29 mmol) ina manner analogous to that described for compound 1E to give a colorlesssyrup (104.9 mg, 89%). HPLC: 3.58 min (retention time) (Conditions: YMCS-5 C-18 (4.6×250 mm), eluting with 0-100% B, 4 min gradient. (A=90%H₂O—10% CH₃CN—0.1% TFA and B=10% H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4mL/min. UV detection at 220 nm. Chiral HPLC: retention time=12.53 min(99.96%); Conditions: AD (4.6×250 mm); Eluted with 20% isopropanol inheptane for 30 min at 1 mL/min. MS (ES) m/z 406 [M+H]⁺.

56F.2-Chloro-4-[(6R,7aS)-6-hydroxy-3-oxotetrahydropyrrolo[1,2-c]imidazol-2(3H)-yl]-3-methylbenzonitrile

The title compound was prepared from compound 56E (95.9 mg, 0.24 mmol)in a manner analogous to that described for compound 1E to give a whitefoam (69.8 mg, 100%). HPLC: 1.66 min (retention time) (Conditions: YMCS-5 C-18 (4.6×250 mm), eluting with 0-100% B, 4 min gradient. (A=90%H₂O—10% CH₃CN—0.1% TFA and B=10% H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4mL/min. UV detection at 220 nm. Chiral HPLC: retention time=15.42 min(100%); Conditions: AD (4.6×250 mm); Eluted with 20% isopropanol inheptane for 30 min at 1 mL/min. MS (ES) m/z 290 [M−H]⁻.

EXAMPLE 57(7S,7aS)-2-Chloro-4-(7-hydroxy-3-oxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To a suspension of(7S,7aR)-2-Chloro-4-(7-hydroxy-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile(23C) (5.00 g, 16.36 mmol) in THF (164 mL) was added LAH (620 mg, 16.34mmol) in two portions. After 30 min, the reaction was quenched by theslow addition of the following: water (0.62 mL) in THF (2 mL), 1.86 mL15% aqueous NaOH, then 1.86 mL water in THF (2 mL). After stirring for15 min, the reaction was filtered through celite, washed with EtOAc, andconcentrated under reduced pressure to obtain 4.45 g of a yellow foam.The foam was taken up in CH₂Cl₂ (142 mL) and cooled to −78° C.Triethylsilane (4.50 mL, 28.17 mmol) and boron trifluoride diethyletherate (3.60 mL, 28.41 mmol) were then added and the cold bath wasremoved. After stirring for 1.5 h, the reaction mixture was poured intosaturated aqueous NaHCO₃ and the layers were separated. The organiclayer was dried (MgSO₄), filtered, concentrated under reduced pressure,and then purified using preparative HPLC (YMC ODS C-18, 30×250 mm,eluting with 50-70% solvent B (A=90% H₂O—10% MeOH—0.1% TFA and B=10%H₂O—90% MeOH—0.1% TFA) over 30 min; Flow rate at 20 mL/min. UV detectionat 220 nm) to provide a white solid (ca. 1 g). The solid was furtherpurified by several recrystallizations (first from MeOH/H₂O then EtOHuntil purity >99% by ¹H NMR) to provide 236 mg of the desired racemicproduct as white needles. A portion of this material (28 mg) was furtherpurified using chiral preparative HPLC (Chiracel OD, 5×50 cm, elutingwith 20% isopropanol/hexane, Flow rate=56 mL/min, UV detection at 220nm) to afford 57 (6.8 mg) (Chiral HPLC: retention time=10.68 min; DaicelChiralcel OD, 4.6×250 mm, eluting with 20% isopropanol in hexane over 30min; Flow rate at 1 mL/min, UV detection at 220 nm; LC/MS m/z 292[M+1]⁺) and 11.5 mg of a white solid which was purified by chiralpreparative HPLC as above followed by preparative HPLC to provide thetitle compound (3.3 mg) (Chiral HPLC: retention time=12.91 min; DaicelChiralcel OD, 4.6×250 mm, eluting with 20% isopropanol/hexane over 30min; Flow rate at 1 mL/min, UV detection at 220 nm). LC/MS m/z 292[M+1]⁺.

EXAMPLE 58(7R,7aR)-Chloro-4-(7-hydroxy-7a-methyl-3-oxotetrahydropyrrolo-[1,2c]imidazol-2-yl)-3-methylbenzonitrile

58A.(2S,3R)-3-(tert-Butyldimethylsilanyloxy)-2-methylpyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester-2-methyl ester

To a solution of (2S,3R)-3-Hydroxy-2-methylpyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester-2-methyl ester (25A) (362 mg, 1.39 mmol) inCH₂Cl₂ (4 mL) was added imidazole (284 mg, 4.17 mmol) thentert-butydimethylsilyl chloride (1.05 g, 6.95 mmol). After stirringovernight, the reaction was partitioned between H₂O and CH₂Cl₂. TheCH₂Cl₂ layer was washed with 1M H₃PO₄ (2×) and brine, dried (MgSO₄),then filtered and concentrated under reduced pressure. The residue waschromatographed (silica gel) eluting with 30% EtOAc/hexane to yield thetitle compound (456 mg) as a white solid.

58B.(2R,3R)-3-(tert-Butyldimethylsilanyloxy)-2-hydroxymethyl-2-methylpyrrolidine-1-carboxylicacid tert-butyl ester

To a solution of(2S,3R)-3-(tert-Butyldimethylsilanyloxy)-2-methylpyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester-2-methyl ester (58A) (1.08 g, 2.90 mmol) in THF(12 mL) at −78° C. was added a 1 M solution of Super-Hydride® in THF(14.50 mL, 14.50 mmol) in three portions over 15 min. After 10 min, thecold bath was removed and the reaction was allowed to warm to rt and wasstirred for 18 h. The reaction was cooled again to −78° C. and moreSuper-Hydride® (7 mL) was added. After stirring an additional 24 h, thereaction was poured into a 1-L Erlenmeyer flask containing ice water andwas then diluted with EtOAc. The layers were separated and the organiclayer washed with 1 M H₃PO₄ (2×), NaHCO₃ and brine, dried (MgSO₄),filtered and concentrated under reduced pressure. The residue wasdiluted with CH₂Cl₂, stirred with silica gel for 10 min, thenconcentrated under reduced pressure and purified via flashchromatography, eluting with 20% EtOAc/hexane to obtain the titlecompound (0.54 g) as a clear oil. MS m/z 346 [M+H]⁺.

58C.(2R,3R)-[3-(tert-Butyldimethylsilanyloxy)-2-methylpyrrolidin-2-yl]methanoltrifluoroacetic acid salt

(2R,3R)-3-(tert-Butyldimethyl-silanyloxy)-2-hydroxymethyl-2-methylpyrrolidine-1-carboxylicacid tert-butyl ester (58B) (269 mg, 0.78 mmol) was stirred in 17%TFA/CH₂Cl₂ (6 mL) for 30 min. The reaction was concentrated to provide abrown oil (334 mg). LC/MS m/z 246 [M+H]⁺

58D.(2R,3R)-3-(tert-Butyldimethylsilanyloxy)-2-hydroxymethyl-2-methylpyrrolidine-1-carboxylicacid (3-chloro-4-cyano-2-methyl-phenyl)-amide

To a solution of(2R,3R)-[3-(tert-Butyldimethylsilanyloxy)-2-methylpyrrolidin-2-yl]methanoltrifluoroacetic acid salt (58C) (167 mg, 0.61 mmol) in CH₂Cl₂ (2.5 mL)at 0° C. were added molecular sieves followed by Hunig's base (0.21 mL,1.22 mL). After stirring for 15 min,2-Chloro-4-isocyanato-3-methylbenzonitrile (23A) was added and the icebath was removed. After 10 min, the reaction was stirred for 2 h andthen diluted with water and CH₂Cl₂. The layers were separated and theorganic layer was washed with brine, dried (MgSO₄), filtered andconcentrated. The resulting solid was purified via preparative HPLC(Luna C-18, 250×21.2 mm, eluting with 60-100% solvent B (A=90% H₂O—10%MeOH—0.1% TFA and B=10% H₂O—90% MeOH—0.1% TFA) over 15 min; Flow rate at10 mL/min; UV detection at 220 nm) to provide the title compound (17 mg)as a white film (LC/MS m/z 438 [M+H]⁺) and di-acylated product (80 mg)as a white solid (LC/MS m/z 630 [M+H]⁺). The di-acylated product (80 mg,0.13 mmol) was dissolved in EtOH (2 mL) and treated with 21% NaOEt (48□L, 0.13 mmol) at rt for 4 h. The reaction was concentrated underreduced pressue then diluted with water and EtOAc. The layers wereseparated, and the aqueous layer acidified with 1 N HCl thenre-extracted with EtOAc. The combined organic layers were washed withbrine, dried (MgSO₄), filtered and concentrated under reduced pressure.The residue was purified via preparative HPLC (Luna C-18, 100×21.2 mm,eluting with 40-100% solvent B (A=90% H₂O—10% MeOH—0.1% TFA and B=10%H₂O—90% MeOH—0.1% TFA) over 10 min; Flow rate at 20 mL/min; UV detectionat 220 nm) to provide the title compound (43 mg) as a white film. LC/MSm/z 438 [M+H]⁺.

58E.(7R,7aR)-4-[7-(tert-Butyldimethylsilanyloxy)-7a-methyl-3-oxotetrahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methyl-benzonitrile

To a solution of(2R,3R)-3-(tert-Butyldimethylsilanyloxy)-2-hydroxymethyl-2-methylpyrrolidine-1-carboxylicacid (3-chloro-4-cyano-2-methyl-phenyl)-amide (58D) (43 mg, 0.10 mmol)in THF (1 mL) at 0° C. was added a 1 M solution of potassiumtert-butoxide in THF (0.24 mL, 0.24 mmol) followed by a solution ofp-toluenesulfonyl chloride (22 mg, 0.12 mmol) in THF (0.5 mL). After 10min, additional potassium tert-butoxide (50 □L) and p-toluenesulfonylchloride (3 mg) were added. After another 10 min, the reaction wasdiluted with water and EtOAc and the layers were separated. The organiclayer was washed with brine, dried (MgSO₄), filtered and concentratedand purified via preparative HPLC (Luna C-18, 100×21.2 mm, eluting with60-100% solvent B (A=90% H₂O—10% MeOH—0.1% TFA and B=10% H₂O—90%MeOH—0.1% TFA) over 12 min; Flow rate at 20 mL/min; UV detection at 220nm) to provide the title compound (17 mg). LC/MS m/z 420 [M+H]⁺.

58F.(7R,7aR)-2-Chloro-4-(7-hydroxy-7a-methyl-3-oxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To a solution of(7R,7aR)-4-[7-(tert-Butyldimethylsilanyloxy)-7a-methyl-3-oxotetrahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile(58E) (17 mg, 0.04 mmol) in THF (2 mL) was added acetic acid (100 □L)and a 1 M solution of TBAF in THF (122 □L, 0.12 mmol). After stirring atrt for 11 h, additional TBAF solution was added (100 □L), and afterstirring for another 6 h, 200 □L TBAF solution was added. The reactionwas stirred overnight, then 100 □L TBAF was added and the reactionstirred an additional 1.5 h. The reaction was then quenched withsaturated aqueous NH₄Cl and extracted with EtOAC. The organic layer waswashed with brine, dried (MgSO₄), filtered, and concentrated underreduced pressure. The resulting residue was purified via preparativeHPLC (Luna C-18, 100×21.2 mm, eluting with 40-100% solvent B (A=90%H₂O—10% MeOH—0.1% TFA and B=10% H₂O—90% MeOH—0.1% TFA) over 10 min; Flowrate at 20 mL/min; UV detection at 220 nm) to provide the title compound(11 mg). LC/MS m/z 306 [M+H]⁺.

EXAMPLE 59(7aR)-2-Chloro-3-methyl-4-(7a-methyl-3,7-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)benzonitrile

To a 2 M CH₂Cl₂ solution of oxalyl chloride (68 □L, 0.13 mmol) in CH₂Cl₂(2 mL) at −78° C. was added DMSO (17□L, 0.25 mmol). After 20 min, asolution of(7R,7aR)-2-chloro-4-(7-hydroxy-7a-methyl-3-oxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile(58F) (17 mg, 0.06 mmol) in CH₂Cl₂ (1 mL) was added. After an additional20 min at −78° C., triethylamine (62 □L, 0.47 mmol) was added, the coldbath was removed, and the reaction mixture was stirred for 20 min. Waterwas added and the layers were separated. The organic layer was washedwith brine then dried (MgSO₄), filtered and concentrated. The resultingresidue was purified via preparative HPLC (YMC ODS C-18, 100×20 mm,eluting with 30-80% solvent B (A=90% H₂O—10% CH₃CN—0.1% TFA and B=10%H₂O—90% CH₃CN—0.1% TFA) over 15 min; Flow rate at 20 mL/min; UVdetection at 220 nm) to provide the title compound (6.3 mg) as a whitesolid. LC/MS m/z 629 [2M+23]⁺.

EXAMPLE 60(7R,7aR)-2-Chloro-4-(7-hydroxy-7a-methyl-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

60A.(2R,3R)-3-(tert-Butyldimethylsilanyloxy)-2-methylpyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester-2-methyl ester

To a solution of(2S,3R)-3-(tert-Butyldimethylsilanyloxy)pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester (51A) (195 mg, 0.54 mmol) in THF(6 mL) at −78° C. was added a 1.8 M solution of LDA (0.66 mL, 1.19mmol). After stirring for 1.5 h at −78° C. and 0.5 h at −30° C., thereaction was cooled again to −78° C. and iodomethane (0.2 mL, 3.21 mmol)was added. The mixture was stirred at −78° C. for 1 h and at −20° C. for4 h. After warming to rt, water and EtOAc were added and the layers wereseparated. The organic layer was washed with brine, dried (Na₂SO₄),filtered then concentrated under reduced pressure. The resulting residuewas purified via preparative HPLC (Luna C-18, 21.1×100 mm, eluting with75-100% solvent B (A=90% H₂O—10% MeOH—0.1% TFA and B=10% H₂O—90%MeOH—0.1% TFA) over 12 min; Flow rate at 20 mL/min. UV detection at 220nm) to provide the title compound (36 mg). LC/MS m/z 374 [M+H]⁺.

60B.(7R,7aR)-4-[7-(tert-Butyldimethylsilanyloxy)-7a-methyl-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methyl-benzonitrile

A solution of(2R,3R)-3-(tert-Butyldimethylsilanyloxy)-2-methyl-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester-2-methyl ester (60A) (52 mg, 0.14 mmol) inCH₂Cl₂ (1.5 mL) and TFA (0.5 mL) was stirred at rt for 2 h. The reactionwas concentrated under reduced pressure and dried under vacuum. Theresidue was dissolved in CH₂Cl₂ (1.5 mL) and Hunig's base (60 μL, 0.35mmol) was added. After stirring at rt for 10 min,2-chloro-4-isocyanato-3-methylbenzonitrile (34 mg, 0.18 mmol) was addedand the reaction was stirred at rt overnight. The reaction was treatedwith DBU (40 μL, 0.27 mmol) and was stirred for 4 h at rt thenconcentrated under reduced pressure. The residue was purified viachromatography (silica gel) eluting with 30% EtOAc in hexane to providethe title compound (54 mg). LC/MS m/z 434 [M+H]⁺.

60C.(7R,7aR)-2-Chloro-4-(7-hydroxy-7a-methyl-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

A solution of(7R,7aR)-4-[7-(tert-Butyldimethylsilanyloxy)-7a-methyl-1,3-dioxotetrahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile(60B) (32 mg, 0.07 mmol) in THF (2 mL) in a plastic vial was cooled to0° C. HF/pyridine complex (0.12 mL) was added and the reaction wasstirred at 0° C. for 1 h and at rt overnight. Saturated aqueous NaHCO₃and CH₂Cl₂ were added. The layers were separated and the organic layerwas concentrated under reduced pressure. The residue was purified viachromatography (silica gel) eluting with 75% EtOAc in hexane to providethe title compound (16 mg). LC/MS m/z 320 [M+H]⁺.

EXAMPLE 614-[(1S,7S,7aR)-7-hydroxy-1-methyl-3-oxo-hexahydro-pyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methyl-benzonitrile

61A.(2R,3S)—N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-hydroxymethylpyrrolidine

Following procedures described for the preparation of(2S,3R)—N-tert-butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)prolinemethyl ester (51A), (2S,3S)-3-hydroxyproline was converted to(2S,3S)—N-tert-butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)prolinemethyl ester. To this material (4.3 g, 12 mmol) in THF (75 mL) at −78°C. under nitrogen was added dropwise 1.0 M lithium triethylborohydridein THF (60 mL, 60 mmol). The reaction was allowed to warm to rt and wasstirred for 4 h. The reaction was quenched by pouring over ice (150 g)and stirring for 30 min. The product was extracted into EtOAc and washedwith saturated aqueous NaHCO₃. The organics were dried over MgSO₄,filtered and concentrated. Purification by flash chromatography (silicagel, 0-15% EtOAc in hexanes) provided the title compound (3.3 g).61B.(2S,3S)—N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-formyl-pyrrolidine

To a solution of 61A (3.3 g, 9.97 mmol) in 10% CH₃CN in CH₂Cl₂ (50 mL)at rt was added NMO (3.0 g, 25.6 mmol) followed by tetrapropyl ammoniumperruthenate (300 mg, 0.85 mmol). After a mild exotherm, the reactionwas stirred at rt for 4 h. The reaction was diluted with hexane (50 mL),mixed well and filtered through a plug of silica gel (˜50 g). The silicaplug was washed liberally with 30% EtOAc in hexane (500 mL). Solventswere removed under vacuum. Purification by flash chromatography (silicagel, 0-15% EtOAc in hexanes) provided the title compound (2.94 g).

61C.(2R,3S)—N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-(1-hydroxyethyl)pyrrolidine

To a solution of intermediate 61B (˜0.05 M in THF) at −78° C. was addeddropwise MeMgBr (3.0 M in Et₂O, 3 equiv). The reaction was stirred at−78° C. for 1.5-2 h (quench small aliquot in 10% MeOH/DCM, spotTLC-silica gel, 30% EtOAc/Hex—stain with 5% HCl/MeOH, heat and thenninhydrin, heat). Quench reaction by addition of HOAc (3 equiv) at −78°C. Warm to rt, dilute with EtOAc and wash with a 1:1 mixture of sat'daq. NaHCO₃ and brine. Back extract aqueous layer. Combine organicextracts, dry over MgSO₄, filter and concentrate. Purification by flashchromatography (silica gel, step gradient 0-10-20% EtOAc in hexane)provided the title compound.

61D.(2S,3S)—N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-(1-hydroxyethyl)pyrrolidine

To intermediate 61C (0.1 M solution in 10% CH₃CN in DCM) add NMO (2equiv) followed by TPAP (0.05 equiv). Provide sufficient ventilation andtake care to watch for exotherm. Stir reaction at rt for 3-5 h. Diluteby half with hexane and allow TPAP to precipitate. Flash filter throughsilica gel plug (20 fold mass of starting alcohol) and elute with 10%EtOAc/Hex until product is not observed in eluent. Removal of solventsgave the title compound. MS (ES) m/z 344.31 [M+H]⁺

61E.(2R,3S)—N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-(1-hydroxyiminoethyl)pyrrolidine

To a solution of 61D (3.86 g, 11.3 mmol) in methanol:water (2:1, 50 mL)was added hydroxylamine HCl salt (3.0 g) followed by pyridine (5.0 mL).The reaction was stirred at rt for 18 h. Extract product into EtOAc,washing once with sat'd aqueous NaHCO₃ and once with water. Back extractthe aqueous washings with EtOAc and combine this with the initialextract. Dry organic extract over MgSO₄, filter and concentrate. Diluteresidue in toluene and remove solvent under vacuum to provide the titlecompound as a white solid (2.9 g). MS (ES) m/z 359.30 [M+H]⁺

61F.(2R,3S)—N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-((1S)-1-aminoethyl)pyrrolidine

To a solution of 61E (3.6 g, 10.0 mmol) in methanol in a pressure vesselwas added Raney Ni in water (˜0.2 g), 10% Pd/C (Degussa type, 200 mg),water (7.5 mL) and ammonia in methanol (2.0 M, 5 mL). The reactionvessel was carefully evacuated under vacuum until the solvent bubbledgently. Hydrogen gas was then introduced to a pressure of 70 psi and thereaction was stirred for 2 min). The reaction vessel was again carefullyevacuated under vacuum until the solvent bubbled gently. Hydrogen gaswas then introduced to a pressure of 70 psi and the reaction was stirredfor 18 h at rt. The reaction was filtered through a pad of celite,taking care to keep the captured catalyst wet with methanol at alltimes. The celite pad was washed with methanol until the product couldno longer be detected in the eluent (TLC, 5% MeOH in CH₂Cl₂, stain withninhydrin and heat). Solvent was removed under vacuum and purificationby flash chromatography (110 g ISCO silica gel cartridge, step gradient0%-5%-10% MeOH in CH₂Cl₂) gave the title compound (2.2 g): MS (ES) m/z345.35 [M+H]⁺

61G. 2-Chloro-4-iodo-3-methylbenzonitrile

To a suspension of CuI (7.5 g, 39.3 mmol) in acetonitrile (150 mL) underN₂ at rt was added tert-butylnitrite (5.7 mL, 47.9 mmol). The reactionmixture was heated to 65° C. for 1 h and then4-amino-2-chloro-3-methyl-benzonitrile (23D) (6.0 g, 36.0 mmol) wasadded and the reaction was heated at 65° C. for 3 h. The reaction wascooled to rt and filtered through a pad of celite. The celite pad waswashed with EtOAc. The organics were washed twice with water, dried overMgSO₄, filtered and concentrated. Purification by flash chromatography(silica gel, 110 g ISCO, 0-5% EtOAc in hexane, step gradient) gave thetitle compound (4.3 g): MS (ES) m/z 278 [M+H]⁺

61H.(2R,3S)-1-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-[(1S)-1-(3-chloro-4-cyano-2-methyl-phenylamino)ethyl]pyrrolidine

To a solution of 61F (325 mg, 0.94 mmol) in degassed toluene:DMSO (2:1,7 mL) at rt was added 61G (260 mg, 0.94 mmol), Cs₂CO₃ (614 mg, 1.89mmol) and a solution of Pd₂(dba)₃ and(S)—N,N-dimethyl-1-[(R)-2-(diphenyphosphino)ferrocenyl]ethylamine (1:6ratio, 0.06 mol %) in nitrogen degassed toluene (3 mL). The reaction wasdegassed with nitrogen for 30 min, sealed and heated at 110° C. for 48h. Cool to rt and extract with EtOAc (50 mL) and wash with waterfollowed by sat'd aqueous NaHCO₃. Dry over MgSO₄, filter andconcentrate. Purification by flash chromatography (40 g ISCO silica gelcartridge, 0-10% EtOAc in hexane gradient) gave the title compound (460mg): MS (ES) m/z 494.29 [M+H]⁺

61I.(2R,3S)-3-(tert-Butyldimethylsilanyloxy)-2-[(1S)-1-(3-chloro-4-cyano-2-methyl-phenylamino)ethyl]pyrrolidine

Intermediate 61H (825 mg, 1.67 mmol) was dried azeotropically withtoluene (2×). The residue was taken up in 15% TFA in CH₂Cl₂ (10 mL) andstirred at rt for 5 h. Toluene (10 mL) was added and solvent was removedunder vacuum. The product was purified by reverse phase HPLC (PhenoenexLuna 30×100 mm S5 C18, 10 min. grad, 25 mL/min, 20-100% B solvent, A=10%MeOH/water+0.1% TFA, B=90% MeOH/Water+0.1% TFA, 4 injections). Solventwas reduced to ˜10% volume and the product was then extracted intoEtOAc, washed with sat. aqueous NaHCO₃, dried over MgSO₄ and filtered.Solvents were removed to provide the title compound (371 mg): MS (ES)m/z 394.53 [M+H]⁺

61J.4-[(1S,7S,7aR)-7-tert-Butyldimethylsilanoxy-1-methyl-3-oxo-hexahydro-pyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of crude 61I (˜1.81 mmol) in CH₂Cl₂ (10 mL) was addedi-Pr₂NEt (2 mL) followed by 20% phosgene in toluene (2.5 mL) dropwise.The reaction was stirred at rt overnight. Reaction was extracted withEtOAc (50 mL) which was washed twice with 1N HCl and then saturatedNaHCO₃. The organic layer was dried over MgSO4, filtered andconcentrated. Purification by flash chromatography (silica gel, 0 to 10%EtOAc in hexane) provided the title compound (369 mg): MS (ES) m/z420.23 [M+H]⁺

61K.4-[(1S,7S,7aR)-7-Hydroxy-1-methyl-3-oxo-hexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of 61J (369 mg) in THF (4 mL) was added TBAF (1.0 M inTHF, 1.3 mL). The reaction was stirred at rt for 1 h. The reaction wasextracted with EtOAc which was washed three times with water, dried overMgSO₄, filtered and concentrated. Purification by reverse phase HPLC(Phenoenex Luna 20×100 mm S5 C18, 10 min. grad, 20 mL/min, 20-100% Bsolvent, A=10% MeOH/water+0.1% TFA, B=90% MeOH/Water+0.1% TFA) providedthe title compound (186 mg): MS (ES) m/z 306.42 [M+H]⁺

EXAMPLE 62 4-[(1R,7R,7aS)-7-Hydroxy-1-methyl-3-oxo-hexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

The title compound was prepared in a fashion similar to Example 61: MS(ES) m/z 306.42 [M+H]⁺

EXAMPLE 634-[(1R,7aS)-1-Methyl-3,7-dioxo-hexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of DMSO (20 μL) in CH₂Cl₂ (1 mL) at −78° C. was addedoxalyl chloride (1.0 M in CH₂Cl₂, 131 μL). This mixture was stirred at−78° C. for 30 min. and then a solution of 61K in CH₂Cl₂ (20 mg in 1mL). The reaction was stirred at −78° C. for 1 h and then i-Pr₂NEt (100μL) was added and the reaction was allowed to warm to rt over 1 h. Thereaction was extracted with EtOAc and washed twice with 1N HCl and oncewith saturated NaHCO₃. The organic layer was dried over MgSO₄, filteredand concentrated. Purification by flash chromatography (silica gel, 40%EtOAc/CH₂Cl₂) provided the title compound (18 mg): MS (ES) m/z 304.42[M+H]⁺

EXAMPLE 64 4-[(1S, 7R,7aR)-7-Hydroxy-1-methyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of 63 (14 mg) in THF (3 mL) at −78° C. was addedK-selectride (1.0 M in THF, 100 μL). The reaction was stirred at −78° C.for 30 min. MeOH (100 μL) was added and the reaction was stirred at −78°C. for an additional 10 min. The reaction was warmed to rt, diluted withEtOAc (25 mL) and washed with saturated NaHCO₃. The organic layer wasdried over MgSO₄, filtered and concentrated. Purification by preperativeHPLC (Phenoenex Luna 20×100 mm S5 C18, 10 min. grad, 20 mL/min, 20-100%B solvent, A=10% MeOH/water+0.1% TFA, B=90% MeOH/Water+0.1% TFA)provided the title compound (8.3 mg): MS (ES) m/z 306.42 [M+H]⁺

EXAMPLE 654-(1R,7aS)-7-Hydroxyimino-1-methyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of 63 (20 mg) in MeOH/water (2:1, 3 mL) was added pyridine(0.5 mL) and NH₂OH.HCl (20 mg). After the reaction was stirred at rtovernight, EtOAc was added and the organic layer was washed once withwater. The organic layer was dried over MgSO₄, filtered andconcentrated. Purification by preperative HPLC (Phenoenex Luna 20×100 mmS5 C18, 10 min. grad, 20 mL/min, 20-100% B solvent, A=10%MeOH/water+0.1% TFA, B=90% MeOH/Water+0.1% TFA) provided the titlecompound (9.1): MS (ES) m/z 319.43 [M+H]⁺

EXAMPLE 66(1S,7R,7aR)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

66A.(2R,3R)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1R)-(2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid tert-butyl ester and66B.(2R,3R)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1S)-(2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid tert-butyl ester

To aldehyde 51C (2.10 g, 6.38 mmol) was addedtrimethyl(trifluoromethyl)silane (800 μL, 6.57 mmol) and cesium fluoride(dried under high vacuum at 130° C. for 12 h) (10.0 mg, 0.0658 mmol).The reaction was stirred at rt for 24 h then heated to 50° C. for 5 h.After cooling to rt, 4 N HCl (ca. 10 mL) was added and the reaction wasstirred overnight. The brown aqueous solution was decanted and the waxyyellow solid was dried under high vacuum overnight then recrystallizedfrom hexane to provide(2R,3R)-3-(tert-butyl-dimethyl-silanyloxy)-2-[(1R)-(2,2,2-trifluoro-1-hydroxy-ethyl)]-pyrrolidine-1-carboxylicacid tert-butyl ester (66A) (1.19 g) as a white solid. The mother liquorwas concentrated and purified via flash chromatography eluting with5-20% ethyl acetate/hexane to provide(2R,3R)-3-(tert-butyl-dimethyl-silanyloxy)-2-[(1S)-(2,2,2-trifluoro-1-hydroxy-ethyl)]-pyrrolidine-1-carboxylicacid tert-butyl ester (66B) (192 mg) as an oil and a mixture of 66A andstarting material 51C (306 mg). The latter mixture was recrystallizedfrom hexane to provide additional 66A (100 mg).

66C.(2R,3R)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1R)-((2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid (3-chloro-4-cyano-2-methylphenyl)amide

To(2R,3R)-3-(tert-butyl-dimethyl-silanyloxy)-2-[(1R)-(2,2,2-trifluoro-1-hydroxy-ethyl)]-pyrrolidine-1-carboxylicacid tert-butyl ester (66A) (1.29 g, 3.23 mmol) in CH₂Cl₂ (24 mL) wasadded trifluoroacetic acid (8 mL). After stirring for 30 min at rt,toluene (ca. 5 mL) was added, the reaction was concentrated underreduced pressure and the brown oil was dried under high vacuumovernight. The resulting brown waxy solid was dissolved in CH₂Cl₂ (32mL) and was cooled to −78° C. DIPEA (1.13 mL, 6.49 mmol) was added andthe whole was stirred for 15 min at −78° C.2-chloro-4-isocyanato-3-methylbenzonitrile (23E) (621 mg, 3.23 mmol) inCH₂Cl₂ (5 mL) was then added and the cold bath was removed. Afterstirring for 2 h (−78° C. to rt), water was added and the layersseparated. The organic layer was washed with brine, dried (MgSO₄),filtered and concentrated. The resulting residue was purified via flashchromatography eluting with 30-75% ethyl acetate/hexane to provide(2R,3R)-3-(tert-butyl-dimethylsilanyloxy)-2-[(1R)-((2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid (3-chloro-4-cyano-2-methyl-phenyl)-amide (1.47 g) as a white solid.LCMS: m/z 492 [M+H]⁺.

66D.(1S,7R,7aR)-2-Chloro-4-(7-tert-butyl-dimethylsilanyloxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 66C (1.44 g, 2.93 mmol) in THF (49 mL) at 0° C. was added a 1M THFsolution of potassium tert-butoxide (7.04 mL, 7.04 mmol) followed byp-toluenesulfonyl chloride (670 mg, 3.52 mmol) in THF (5 mL). The coldbath was removed and additional p-toluenesulfonyl chloride was addeduntil starting material was consumed (ca. 50 mg). The reaction wasstirred at rt for 1.5 h then heated to 60° C. After 3 h, additional 1 Mpotassium tert-butoxide (1.00 mL) was added and the was reaction heatedfor an additional hour. The whole was cooled to rt and was diluted withwater and EtOAc. The layers were separated and the organic layer washedwith brine. The aqueous layer was acidified to pH 1 with 1 N HCl and wasextracted with ethyl acetate. The organic layers were combined and dried(MgSO₄), filtered and concentrated. The residue was purified via flashchromatography eluting with 30-75% ethyl acetate/hexane to provide thetitle compound (956 mg) as a yellow foam.

66E.(1S,7R,7aR)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 66D (956 mg, 2.02 mmol) in THF (20 mL) was added a 1 M THF solutionof TBAF (2.02 mL, 2.02 mmol). After stirring at rt for 1 h, additionalTBAF was added (500 μL) and the reaction was stirred for 45 min.Saturated aqueous ammonium chloride and EtOAc were added and the layerswere separated. The organic layer was washed with brine then dried(MgSO₄), filtered and concentrated. The resulting residue was purifiedvia preparative HPLC (YMC ODS C-18, 30×250 mm, eluting with 50-80%solvent B (A=90% H₂O—10% MeOH and B=10% H₂O—90% MeOH) over 15 min; Flowrate at 20 mL/min; UV detection at 220 nm) to provide the title compound(617 mg) as a white solid. MS: m/z 358 [M−H]⁻.

EXAMPLE 67(1R,7R,7aR)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxo-hexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

The title compound was prepared from(2R,3R)-3-(tert-butyl-dimethyl-silanyloxy)-2-[(1S)-(2,2,2-trifluoro-1-hydroxy-ethyl)]-pyrrolidine-1-carboxylicacid tert-butyl ester (66B) following procedures analogous to thosefound in Example 66. MS: m/z 358 [M−H]⁻

EXAMPLE 68(1S,7aR)-2-Chloro-4-(7-hydroxyimino-1-trifluoromethyl-3-oxohexahydropyrrolo[2-c]imidazol-2-yl)-3-methylbenzonitrile

68A.(1S,7aR)-2-Chloro-4-(1-trifluoromethyl-3,7-dioxo-hexahydro-pyrrolo[1,2-c]imidazol-2-yl)-3-methyl-benzonitrile

To 66E (100 mg, 0.279 mmol) in CH₂Cl₂ was added Dess-Martin periodinane(236 mg, 0.556 mmol) and the reaction was stirred at rt overnight.Saturated aqueous Na₂S₂O₃ and NaHCO₃ were added and the whole wasstirred vigorously for 0.5 h. The layers were separated, the organiclayer was washed with a mixture of saturated aqueous Na₂S₂O₃ and NaHCO₃followed by brine, dried (MgSO₄), and was then filtered and concentratedto obtain the title compound (96.0 mg) as a white film.

68B.(1S,7aR)-2-Chloro-4-(7-hydroxyimino-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 68A (96.0 mg, 0.269 mmol) in MeOH (2.7 mL) and water (5 drops) wasadded pyridine (109 μL) and hydroxylamine hydrochloride (93.0 mg, 1.34mmol) and the whole was stirred overnight at rt. The reaction wasconcentrated then diluted with ethyl acetate and washed with water andbrine. The organic layer was dried (MgSO₄), filtered and concentrated.The resulting residue was purified via preparative HPLC (YMC ODS C-18,30×100 mm, eluting with 60-100% solvent B (A=90% H₂O—10% MeOH and B=10%H₂O—90% MeOH) over 10 min; Flow rate at 40 mL/min; UV detection at 220nm) to provide a solid which was re-purified via preparative HPLC (YMCODS C-18, 30×100 mm, eluting with 40-70% solvent B (A=90% H₂O—10% CH₃CNand B=10% H₂O—90% CH₃CN) over 12 min; Flow rate at 40 mL/min; UVdetection at 220 nm) to provide the title compound (48.0 mg) as a whitesolid. MS: m/z 371 [M−H]⁻.

EXAMPLE 69(1S,7S,7aS)-2-Chloro-4-(7-amino-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

69A.(1S,7R,7aR)-2-Chloro-4-(7-methanesulfonoxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 66E (54.0 mg, 0.150 mmol) in pyridine (1 mL) was added methansulfonylchloride (14.0 μL, 0.182 mmol). After stirring at rt for 3 h, additionalmethansulfonyl chloride (14.0 μL) was added. The reaction was heated to65° C. and after 5 h, additional methansulfonyl chloride (14.0 μL) wasadded. After stirring at 65° C. overnight, the reaction was heated to80° C. and additional methansulfonyl chloride (20.0 μL) was added. After2 h, the reaction was cooled to rt and was diluted with water and EtOAc.The layers were separated and the organic layer was washed with 1 N HCl(3×), then dried (MgSO₄), filtered and concentrated. The resultingresidue was purified via preparative HPLC (YMC ODS C-18, 21×100 mm,eluting with 50-85% solvent B (A=90% H₂O—10% MeOH and B=10% H₂O—90%MeOH) over 15 min; Flow rate at 20 mL/min; UV detection at 220 nm) toprovide starting material (22.0 mg) and the title compound 69A (14.0mg). The recovered starting material was diluted with pyridine (2 mL)and treated with methanesulfonyl chloride (50.0 μL) at rt overnight toprovide, after work-up and purification as above, additional 69A (14.0mg).

69B.(1S,7S,7aR)-2-Chloro-4-(7-azido-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 69A in DMF (1 mL) was added sodium azide (17.0 mg, 0.262 mmol) andthe reaction was stirred overnight at 75° C. The reaction was cooled tort then diluted with water and EtOAc. The layers were separated and theorganic layer was washed with water (3×), then dried (MgSO₄), filteredand concentrated. The resulting residue was purified via flashchromatography eluting with 50% EtOAc/hexane to provide the titlecompound (15.0 mg) as a clear oil.

69C. (1S,7S,7aS)-2-Chloro-4-(7-amino-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 69B (15.0 mg, 0.0391 mmol) in EtOH (1 mL) was added platinum (IV)oxide (5 mg) and hydrogen was bubbled through the system for 10 min. Thehydrogen outlet was removed and the system was stirred under a hydrogenatmosphere. After 2h, the reaction was filtered through celite andconcentrated. The resulting residue was purified via preparative HPLC(YMC ODS C-18, 21×100 mm, eluting with 50-100% solvent B (A=90% H₂O—10%MeOH and B=10% H₂O—90% MeOH) over 10 min; Flow rate at 40 mL/min; UVdetection at 220 nm) to provide the title compound (6.00 mg) as a clearfilm. MS: m/z 419 [M+HCO₃]⁻

EXAMPLE 70 (1S,7R,7aS)- and(1S,7S,7aS)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

70A. (1S,7aS)- and(1S,7aR)-2-Chloro-4-(1-trifluoromethyl-3,7-dioxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To a 2M CH₂Cl₂ solution of oxalyl chloride (167 μL, 0.334 mmol) inCH₂Cl₂ (3 mL) at −78° C. was added DMSO (43 μL, 0.555 mmol). Afterstirring at the same temperature for 20 min, a solution of 66E (89.0 mg,0.248 mmol) in CH₂Cl₂ (1 mL) was added. After an additional 20 min at−78° C., triethylamine (156 μL, 1.12 mmol) was added then the reactionwas lifted out of the cold bath and stirred for 1 h. Water was added andthe were layers separated. The organic layer was washed with brine thendried (MgSO₄), filtered and concentrated. The resulting residue waspurified via flash chromatography eluting with 30% EtOAc/hexane toprovide a 3.3:1 mixture (major and minor products not assigned) of(1S,7aS)-and(1S,7aR)-2-chloro-4-(1-trifluoromethyl-3,7-dioxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile(74.0 mg).

70B.(1S,7R,7aS)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methyl-benzonitrileand

70C.(1S,7S,7aS)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

A mixture of (1S,7aS)- and(1S,7aR)-2-chloro-4-(1-trifluoromethyl-3,7-dioxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile(70A) (74.0 mg, 0.218 mmol) in EtOAc (2 mL) was hydrogenated in thepresence of platinum (IV) oxide (5 mg) for 3 h. The reaction wasfiltered through celite, concentrated and purified via preparative HPLC(YMC ODS C-18, 21×100 mm, eluting with 40-75% solvent B (A=90% H₂O—10%CH₃CN and B=10% H₂O—90% CH₃CN) over 10 min; Flow rate at 40 mL/min; UVdetection at 220 nm) to provide two products. To obtain analyticallypure samples, the two products were purified two more times viapreparative HPLC (YMC ODS C-18, 21×100 mm, first eluting with 40-70%solvent B (A=90% H₂O—10% CH₃CN and B=10% H₂O—90% CH₃CN) over 10 min;Flow rate at 40 mL/min; UV detection at 220 nm), then using MeOH/waterunder similar conditions to provide(1S,7R,7aS)-2-chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxo-hexahydro-pyrrolo[1,2-c]imidazol-2-yl)-3-methyl-benzonitrile(70B) (11.0 mg, MS: m/z 717 [2M−H]⁻) and(1S,7S,7aS)-2-chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile(70C) (4.00 mg, MS: m/z 717 [2M−H]⁻).

EXAMPLE 71(1R,7R,7aR)-2-Chloro-4-(7-hydroxy-1-isopropyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

71A.(2R,3R)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1S)-(1-hydroxy-2-methylpropyl)]pyrrolidine-1-carboxylicacid tert-butyl ester

To 51C (6.50 g, 19.8 mmol) in THF (100 mL) at −78° C. was added a 2M THFsolution of isopropylmagnesium chloride (30.0 mL, 60.0 mmol) over 20min. The cold bath was removed and the reaction was stirred for 1 h. Thereaction was then cooled to −60° C. and was quenched with saturatedaqueous ammonium chloride. After warming to rt, the whole was dilutedwith EtOAc. The layers were separated and the organic layer washed withwater and brine, then dried (MgSO₄), filtered and concentrated. Theresulting residue was purified via flash chromatography eluting with 20%EtOAc/hexane to provide the title compound (5.90 g).

71B.(2R,3R)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1S)-(1-hydroxy-2-methylpropyl]pyrrolidine-1-carboxylicacid (3-chloro-4-cyano-2-methylphenyl)amide

(2R,3R)-3-(tert-butyl-dimethylsilanyloxy)-2-[(1S)-(1-hydroxy-2-methylpropyl]pyrrolidine-1-carboxylicacid (3-chloro-4-cyano-2-methylphenyl)amide (71B) (5.90 g) was preparedfrom 71A (5.90 g) following procedures analogous to those found inExample 66 for the preparation of 66C.

71C.(1R,7R,7aR)-2-Chloro-4-(7-tert-butyl-dimethylsilanyloxy-1-isopropyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 71B (3.00 g, 6.45 mmol) in THF (80 mL) at 0° C. was added a 1M THFsolution of LiHMDS (15.5 mL, 15.5 mmol) followed by p-toluenesulfonylchloride (1.84 g, 9.65 mmol) in THF (27 mL). The cold bath was removedand the reaction was stirred for 1 h. The reaction was diluted withwater and EtOAc. The layers were separated and the organic layer washedwith brine. The aqueous layer was acidified to pH 1 with 1 N HCl and wasextracted with EtOAc. The organic layers were combined and dried(MgSO₄), filtered and concentrated. The residue was purified via flashchromatography eluting with 30-75% EtOAc/hexane to provide the titlecompound (2.25 g) as a light yellow solid. LCMS: m/z 448 [M+H]⁺.

71D.(1R,7R,7aR)-2-Chloro-4-(7-hydroxy-1-isopropyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 71C (2.00 g, 4.47 mmol) in THF (45 mL) was added a 1 M THF solutionof TBAF (4.47 mL, 4.47 mmol). After stirring at rt for 45 min, saturatedaqueous ammonium chloride and EtOAc were added and the layers wereseparated. The organic layer was washed with brine then dried (MgSO₄),filtered and concentrated. The resulting residue was purified via flashchromatography eluting with 75-100% EtOAc/hexane to provide the titlecompound (1.36 g) which contained a minor impurity (6.76%). To obtain ananalytically pure sample, a portion of the product (ca. 250 mg) waspurified via preparative HPLC (YMC ODS C-18, 30×100 mm, eluting with 25%solvent B (solvent B=10% H₂O—90% CH₃CN—0.1% TFA) for 25 min; Flow rateat 40 mL/min; UV detection at 220 nm) to provide, afterrecrystallization from MeOH/water, the title compound (106 mg) as whiteneedles. MS: m/z 665 [2M−H]⁻

EXAMPLE 72(1S,7R,7aR)-2-Fluoro-4-(7-hydroxy-1-trifluoromethyl-3-oxo-hexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

72A. N-(3-Fluoro-2-methylphenyl)acetamide

To a solution of 3-fluoro-2-methylaniline (2.65 g, 21.2 mmol) in EtOH(25 mL) at 0° C. was added acetic anhydride (2.60 g, 25.5 mmol). Themixture was stirred at rt overnight then concentrated under reducedpressure to give the title compound (3.54 g). LCMS: m/z 168 [M+H]⁺

72B. N-(4-Bromo-3-fluoro-2-methylphenyl)acetamide

To a solution of N-(3-fluoro-2-methylphenyl)acetamide (72A) (1.71 g,10.2 mmol) in acetic acid (15 mL) at 0° C. was added bromine (1.68 mL,32.8 mmol) over 15 min. The mixture was stirred at rt for 2 h, and wasthen poured into ice (50 g). The mixture was filtered and concentratedunder reduced pressure to give the title compound (2.26 g). LCMS: m/z246 [M+H]⁺

72C. N-(4-Cyano-3-fluoro-2-methylphenyl)acetamide

To a solution of N-(4-bromo-3-fluoro-2-methylphenyl)acetamide (72B)(2.18 g, 8.86 mmol) in DMF (20 mL) was added CuCN (2.61 g, 29.1 mmol).The mixture was stirred at 150° C. for 12 h and was then poured into ice(50 g). After warming to rt, the product was extracted with EtOAc (3×50mL). The combined organic layers were washed with brine, dried andconcentrated. The residue was purified via flash chromatography elutingwith a solution of 5% methanol in 95% of EtOAc/hexane (1:1) to providethe title compound (1.51 g) as a white solid. LCMS: m/z 193 [M+H]⁺

72D. 4-Amino-2-fluoro-3-methylbenzonitrile

N-(4-cyano-3-fluoro-2-methyl-phenyl)-acetamide (72C) (450 mg, 2.34 mmol)was dissolved in 1:1 conc. HCl/EtOH (20 mL,) and the mixture wasrefluxed for 1 h. The reaction was cooled to rt and concentrated. Theresidue was dissolved in EtOAc (30 mL), washed with sat. aq. NaHCO₃ andbrine, dried (MgSO₄), filtered and concentrated. The residue wasrecrystallized from EtOAc/Hexane to provide the title compound (340 mg).LC/MS: m/z 151 [M+H]⁺

72E. 2-Fluoro-4-isocyanato-3-methylbenzonitrile

2-Fluoro-4-isocyanato-3-methylbenzonitrile was prepared from4-amino-2-fluoro-3-methyl-benzonitrile (72D) following a procedureanalogous to the procedure used in the preparation of compound 2E(Example 2).

72F.(2R,3R)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1R)-((2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid (3-fluoro-4-cyano-2-methylphenyl)amide

To(2R,3R)-3-(tert-butyl-dimethylsilanyloxy)-2-[(1R)-(2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid tert-butyl ester (66A) (233 mg, 0.584 mmol) in CH₂Cl₂ (4 mL) wasadded trifluoroacetic acid (1 mL). The mixture was stirred for 30 min atrt, concentrated under reduced pressure, azeotroped from toluene, thendried under high vacuum for 3 h. The resulting brown waxy solid wasdissolved in CH₂Cl₂ (5 mL) and DIPEA (0.300 mL, 1.72 mmol) followed by2-fluoro-4-isocyanato-3-methylbenzonitrile (72E) (133 mg, 0.693 mmol)were added. The mixture was stirred at rt overnight and then wasfiltered and concentrated. The resulting residue was purified by flashchromatography eluting with 0-40% EtOAc/hexane to provide the titlecompound (533 mg) as a white solid. LCMS: m/z 476 [M+H]⁺.

72G.(2S,3R)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1R)-((2,2,2-trifluoro-1-methanesulfonoxy)]pyrrolidine-1-carboxylicacid (3-fluoro-4-cyano-2-methylphenyl)amide

To 72F (100 mg, 0.211 mmol) in CH₂Cl₂ (3 mL) at 0° C. was added a 1M THFsolution of potassium tert-butoxide (0.45 mL, 1 mmol), followed bymethanesulfonyl chloride (76 mg, 0.663 mmol). The mixture was stirred atrt overnight. The reaction was concentrated and the residue was purifiedvia flash chromatography eluting with 0-20% EtOAc/hexane to provide thetitle compound (117 mg) as a white solid. LCMS: m/z 554 [M+H]⁺

72H.(1S,7R,7aR)-2-Fluoro-4-(7-tert-butyl-dimethylsilanyloxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 72G (117 mg, 0.211 mmol) in MeOH (3mL) at 0° C. was added a 25% (wt)MeOH solution of NaOMe (89.0 mg, 0.412 mmol) and the mixture wasrefluxed 1 h. After cooling to rt and concentrating, the residue waspurified via preparative HPLC (YMC ODS C-18, 30×250 mm, eluting with50-100% solvent B (A=90% H₂O—10% MeOH and B=10% H₂O—90% MeOH) over 15min; Flow rate at 20 mL/min; UV detection at 220 nm) to provide thetitle compound (17.0 mg) as a white solid. LCMS: m/z 458 [M+H]⁺

72I.(1S,7R,7aR)-2-Fluoro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 72H (17.0 mg, 0.0371 mmol ) in THF (2mL) was added a 1 M THF solutionof TBAF (0.230 mL, 0.230 mmol). After stirring at rt for 1 h, saturatedaqueous ammonium chloride and EtOAc were added and the layers wereseparated. The organic layer was washed with brine then dried (MgSO₄),filtered and concentrated. The resulting residue was purified via flashchromatography eluting with 0-75% EtOAc/hexane to provide the titlecompound (12.3 mg) as a white solid. LCMS: m/z 344 [M+H]⁺

EXAMPLE 73(1S,7R,7aR)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxo-hexahydropyrrolo[1,2-c]imidazol-2-yl)-3-fluorobenzonitrile

73A. 4-Amino-2-chloro-3-fluorobenzaldehyde

To 3-chloro-2-fluoroaniline (10.0 g, 68.7 mmol) in DMSO (500 mL) wasadded copper (II) chloride (18.5 g, 137.4 mmol) and conc. HCl (50 mL).The whole was heated to 90° C. for 13 h. The reaction was cooled to 0°C. and 4N NaOH was added dropwise to adjust to pH 8. The reaction wasdiluted with water and extracted with Et₂O/EtOAc (1:1). The organiclayer was washed with brine, and was dried, filtered and concentrated.The resulting residue was purified via silica gel chromatography elutingwith 5-30% EtOAc/hexane to provide 4-amino-2-chloro-3-fluorobenzaldehyde(1.00 g) as a yellow powder.

73B. 4-Amino-3-fluoro-2-chlorobenzonitrile

To a solution of hydroxylamine hydrochloride (336 mg, 4.84 mmol) inwater (1.2 mL) was added 4-amino-2-chloro-3-fluorobenzaldehyde (73A)(0.800 g, 4.61 mmol) and pyridine (2.5 mL). After stirring at rt for 1h, copper (II) sulfate pentahydrate (230 mg, 0.922 mmol) was addedfollowed by a solution of triethylamine (1.40 mL, 9.68 mmol) in CH₂Cl₂(2.5 mL). To the resulting dark green reaction mixture was then added asolution of DCC (1.14 g, 5.53 mmol) in CH₂Cl₂ (10 mL) and the reactionwas stirred for 2 h. Formic acid (1 mL) was added and the reaction wasstirred for 20 min, filtered through celite, and concentrated. Theresulting residue was purified via silica gel chromatography elutingwith 20-30% EtOAc/hexane to provide the title compound (0.780 g) as alight brown solid.

73C. 4-Isocyanato-3-fluoro-2-chlorobenzonitrile

4-Isocyanato-3-fluoro-2-chlorobenzonitrile was prepared from4-amino-3-fluoro-2-chlorobenzonitrile (73B) in a manner similar to thatdescribed in Example 2 for the preparation of 2E.

73D.(1S,7R,7aR)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydro-pyrrolo[1,2-c]imidazol-2-yl)-3-fluorobenzonitrile

The title compound was prepared from(2R,3R)-3-(tert-butyl-dimethylsilanyloxy)-2-[(1R)-(2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid tert-butyl ester (66A) and2-chloro-4-isocyanato-3-fluorobenzonitrile (73C) following proceduresanalogous to those found in Example 72. LCMS: m/z 364 [M+H]⁺

EXAMPLE 74(1S,7R,7aR)-2-Trifluoromethyl-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

The title compound was prepared from(2R,3R)-3-(tert-butyl-dimethylsilanyloxy)-2-[(1R)-(2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid tert-butyl ester (66A) and isocyanate 18E following proceduresanalogous to those found in Example 72. LCMS: m/z 394 [M+H]⁺

EXAMPLE 75(1S,7S,7aR)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

75A.(2S,3S)-3-(tert-Butyldimethylsilanyloxy)-2-formylpyrrolidine-1-carboxylicacid tert-butyl ester

To (2R,3S)-3-(tert-butyl-dimethylsilanyloxy)-2-hydroxymethylpyrrolidine-1-carboxylic acid tert-butyl ester (61A) (9.85 g, 29.7 mmol)in CH₂Cl₂ (200 mL) at 0° C. was added Dess-Martin periodinane. The icebath was removed and the reaction was warmed to rt. After 2 h, saturatedaqueous Na₂S₂O₃ and NaHCO₃ (ca. 100 mL each) were added and the reactionmixture was stirred vigorously for 0.5 h. The layers were separated, andthe organic layer was washed with a mixture of saturated aqueous Na₂S₂O₃and saturated aqueous NaHCO₃ followed by brine, dried (MgSO₄), and wasthen filtered and concentrated to obtain the title compound (9.23 g) asa yellow oil.

75B.(2R,3S)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1R)-(2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid tert-butyl ester and

75C.(2R,3S)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1S)-(2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid tert-butyl ester

To 75A (1.00 g, 3.04 mmol) was added trimethyl(trifluoromethyl)silane(550 mg, 3.87 mmol) and cesium fluoride (10 mg, dried under high vacuumat 130° C. for 12 h). The mixture was stirred at rt for 24 h, and thenwas heated to 50° C. for 5 h. After cooling to rt, 4N HCl (ca. 10 mL)was added and the reaction was stirred overnight. The product wasextracted with EtOAc (3×30 mL). The organic layer was washed withsaturated aqueous NaHCO₃ and brine, then dried (MgSO₄), filtered andconcentrated. The residue was purified via flash chromatography elutingwith 0-15% ethyl acetate/hexane to provide(2R,3S)-3-(tert-butyl-dimethyl-silanyloxy)-2-[(1R)-(2,2,2-trifluoro-1-hydroxy-ethyl)]-pyrrolidine-1-carboxylicacid tert-butyl ester (75B) (357 mg) as a colorless oil and(2R,3S)-3-(tert-butyl-dimethylsilanyloxy)-2-[(1S)-(2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid tert-butyl ester (75C) (380 mg) as a white solid.

75D.(2R,3S)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1R)-((2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid (3-chloro-4-cyano-2-methylphenyl)amide

To 75B (490 mg, 1.25 mmol) in CH₂Cl₂ (10 mL) was added trifluoroaceticacid (3 mL). After stirring for 30 min at rt, the reaction wasconcentrated under reduced pressure, azeotroped from toluene and driedunder high vacuum for 3 h. The resulting brown waxy solid was dissolvedin CH₂Cl₂ (5 mL). DIPEA (0.65 mL, 3.73 mmol) was added, followed by2-chloro-4-isocyanato-3-methylbenzonitrile (23E) (285 mg, 1.48 mmol).The mixture was stirred at rt overnight and was then filtered andconcentrated. The resulting residue was purified via flashchromatography eluting with 0-40% EtOAc/hexane to provide the titlecompound (533 mg) as a white solid. LCMS: m/z 492 [M+H]⁺.

75E.(2S,3S)-3-(tert-Butyl-dimethylsilanyloxy)-2-[(1R)-((2,2,2-trifluoro-1-p-toluenesulfonoxy)]pyrrolidine-1-carboxylicacid (3-chloro-4-cyano-2-methylphenyl)amide

To 75D (280 mg, 0.570 mmol) in THF (3 mL) at 0° C. was added a 1 M THFsolution of potassium tert-butoxide (1.14 mL, 1.14 mmol) and tosylchloride (380 mg, 1.99 mmol) and the mixture was stirred at rtovernight. The reaction was diluted with CH₂Cl₂ (30 mL), washed with 1MH₃PO₄, sat'd NaHCO₃ and brine, dried (MgSO₄), filtered and concentrated.The resulting residue was purified via flash chromatography eluting with0-40% EtOAc/hexane to provide the title compound. m/z 646 [M+H]⁺.

75F.(1S,7S,7aR)-2-Chloro-4-(7-tert-butyl-dimethylsilanyloxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To 75E (50.0 mg, 0.0775 mmol) in 1,4-dioxane (2 mL) at 0° C. was added a1M THF solution of potassium tert-butoxide (0.2 mL). The mixture washeated to 170° C. in a microwave oven for 15 min. The reaction wasconcentrated and purified via preparative HPLC (YMC ODS C-18, 30×250 mm,eluting with 50-100% solvent B (A=90% H₂O—10% MeOH and B=10% H₂O—90%MeOH) over 15 min; Flow rate at 20 mL/min; UV detection at 220 nm) toprovide the title compound (7.20 mg). m/z 474 [M+H]⁺.

75G.(1S,7S,7aR)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

To a solution of 75F (8.20 mg, 0.173 mmol) in THF (2 mL) was added a 1 MTHF solution of TBAF (0.120 mL, 0.120 mmol). After stirring at rt for 1h, saturated aqueous ammonium chloride and EtOAc were added and thelayers were separated. The organic layer was washed with brine thendried (MgSO₄), filtered and concentrated. The resulting residue waspurified via preparative HPLC (YMC ODS C-18, 30×250 mm, eluting with50-90% solvent B (A=90% H₂O—10% MeOH and B=10% H₂O—90% MeOH) over 15min; Flow rate at 20 mL/min; UV detection at 220 nm) to provide thetitle compound (4.10 mg) as a white solid. MS: m/z 360 [M+H]^(+.)

EXAMPLE 76(1R,7S,7aR)-2-Chloro-4-(7-hydroxy-1-trifluoromethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile

The title compound was prepared from(2R,3S)-3-(tert-butyl-dimethylsilanyloxy)-2-[(1S)-(2,2,2-trifluoro-1-hydroxyethyl)]pyrrolidine-1-carboxylicacid tert-butyl ester (75C) following procedures analogous to thosefound in Example 75. MS: m/z 360 [M+H]⁺

EXAMPLE 774-[(8R,8aR)-8-Hydroxy-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrileand EXAMPLE 784-[(8S,8aS)-8-Hydroxy-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile

77A. (±)-(cis)-3-Hydroxypipecolic acid

A mixture of 3-hydroxy picolinic acid (20 g, 0.14 mol), 10% rhodiumhydroxide on carbon (The catalyst was prepared according to theliterature procedure: William M. Pealman, Tetrahedron Letters, 1967, 8,1663-1664) (13.2 g), 20% palladium hydroxide on carbon (4.0 g) in 30%ammonium hydroxide solution (40 mL) and water (280 mL) were stirredunder hydrogen at 75 psi for 6 days. The catalysts were removed byfiltration and the filtrate was concentrated to yield the crude product77A as a white foam (18.93 g): MS (ES): m/z 146 [M+1]⁺

77B. (±)-(2S,3R)-3-Hydroxypipecolic acid methyl ester

A solution of 3-hydroxy-pipecolic acid 77A (crude, 21.7g, 0.15 mol) inMeOH (420 mL) was saturated with HCl gas and heated at reflux overnight.Removal of solvent and coevaporation of the residue with diethyl ethergave the hydrochloric acid salt of the title compound 77B as a yellowfoam (24.9 g).

77C. (±)-(2S,3R)-1-tert-Butoxycarbonyl-3-hydroxypipecolic acid methylester

To a mixture of 3-hydroxy-pipecolic acid methyl ester 77B (10.5 g, 66mmol) and TEA (27.6 mL, 198 mmol, 3.0 equiv) in DCM (150 mL) was addedBoc₂O (21.6 g, 99 mmol, 1.5 equiv). The mixture was stirred at rtovernight. The mixture was concentrated and the residue was diluted withEtOAc (200 mL), washed with 0.5 N HCl (3×30 mL), saturated sodiumbicarbonate solution (2×30 mL), brine (2×30 mL), dried (MgSO₄) andevaporated to give the crude product. Purification of the crude productby flash chromatography (0-100% EtOAc/hexane) yield the desired product77C (5.5 g) as a racemic mixture: HPLC (Phenomenex Luna 5μ C18 4.6×50mm, linear gradient over 4 min) retention time 2.48 min (100%).

77D.(±)-(2S,3R)-1-tert-Butoxycarbonyl-3-(tert-butyldimethylsilanyloxy)pipecolicacid methyl ester

To a solution of 77C (3.3 g, 12.7 mmol) in DCM (50 mL) was added DIPEA(6.63 mL, 38.1 mmol), tert-butyldimethylsilyl chloride (3.83 g, 25.4mmol, 2.0 equiv) and DMAP (310 mg, 5.24 mmol, 0.2 equiv). The resultingmixture was stirred at rt overnight. TLC showed no completion andtherefore another 2.2 mL of DIPEA and 1.91 g of tert-butyldimethylsilylchloride were added and stirred at rt for 3 days. The mixture wasconcentrated and the residue was diluted with EtOAc (100 mL), washedwith 0.5 N HCl (2×30 mL), brine (2×30 mL), dried (MgSO₄) and evaporatedto give the crude product as a light pink oil. Purification of the crudeproduct by flash chromatography (5-10% EtOAc/hexane) gave the desiredproduct 77D (3.8 g, 81%): R_(f) (10% EtOAc/hexane) 0.25; HPLC(Phenomenex Luna 5 u C18 4.6×50 mm, linear gradient over 4 min)retention time 4.3 min (100%); MS (ES) m/z 374 [M+1]⁺.

77E.(±)-(2S,3R)-1-tert-Butoxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-hydroxymethylpiperidine

To a solution of 77D (1.0 g, 2.7 mmol) in THF (10 mL) at −78° C. wasadded a solution of LiEt₃BH (1.0 M in THF, 13.5 mL, 13.5 mmol, 5 equiv).The mixture was warmed to rt and stirred at rt for 4 h. The mixture waspoured over ice and stirred vigorously for 15 min. The mixture wasextracted with EtOAc (2×50 mL). The combined organic layers were washedwith 0.5 N HCl (20 mL), saturated sodium bicarbonate solution (20 mL),brine (20 mL), dried (MgSO₄) and evaporated to give the crude product asa colorless oil. Purification of the crude product by flashchromatography (5-50% EtOAc/hexane) gave the desired product 77E (960mg): R_(f) (50% EtOAc/hexane) 0.64; HPLC (Phenomenex Luna 5 u C18 4.6×50mm, linear gradient over 4 min) retention time 4.1 min (100%); MS (ES)m/z 346.1 [M+1]⁺; HRMS calcd for C₁₇H₃₆NO₄Si 346.2414, Found 346.2419.

77F.(±)-(2S,3R)-1-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-formylpiperidine

To a solution of 77E (800 mg, 2.3 mmol) in DCM (15 mL) was added4-methylmorpholine N-oxide (541 mg, 4.6 mmol, 2 equiv) followed by theaddition of TPAP (177 mg, 0.5 mmol, 0.22 equiv). The mixture was stirredat rt for 3 h. The mixture was diluted with hexane (10 mL), loaded on alayer of silica gel and eluted with 25% EtOAc/hexane. The filtrate wasconcentrated to gave the desired product 77F (711 mg, 90%): R_(f) (30%EtOAc/hexane) 0.87; HPLC (Phenomenex Luna 5 u C18 4.6×50 mm, lineargradient over 4 min) retention time 4.1 min (100%); MS (ES) m/z 344.1[M+1]⁺; HRMS calcd for C₁₇H₃₄NO₄Si 344.2257, Found 344.2259.

77G.(±)-(2R,3R)-1-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-[(3-chloro-4-cyano-2-methylphenylamino)-methyl]piperidine

A mixture of 77F (660 mg, 1.92 mmol),4-amino-2-chloro-3-methylbenzonitrile (384 mg, 2.3 mmol, 1.2 equiv) andHOAc (165 μL, 2.88 mmol, 1.5 equiv) in 1,2-DCE (10 mL) was stirred at rtfor 30 min and Na(OAc)₃BH (610 mg, 2.88 mmol, 1.5 equiv) was added. Thereaction was stirred at rt overnight. Another 407 mg of Na(OAc)₃BH (610mg, 1.92 mmol, 1.0 equiv) was added and stirred at rt for another 24 h.The mixture was concentrated and the residue was diluted with EtOAc (30mL), washed with saturated sodium bicarbonate solution (2×15 mL), brine(2×15 mL), dried (MgSO₄) and evaporated to give the crude product.Purification of the crude product by flash chromatography (0-50%EtOAc/hexane) yield the desired product 77G (192 mg, 20%) as a whitefoam: R_(f) (30% EtOAc/hexane) 0.37; HPLC (Phenomenex Luna 5μ C18 4.6×50mm, linear gradient over 4 min) retention time 4.6 min (100%); MS (ES)m/z 344.1 [M+1]⁺; HRMS calcd for C₂₅H₄₁N₃O₃SiCl 494.2606, Found494.2613.

77H.(±)-(2S,3R)-3-(tert-Butyldimethylsilanyloxy)-2-[(3-chloro-4-cyano-2-methylphenylamino)methyl]piperidine

A solution of 77G (170mg, 0.34 mmol) in 5% TFA/DCM (5 mL) was stirred atrt for 5 h and LC-MS showed completion. The mixture was concentrated andthe residue was dissolved in EtOAc (30 mL), washed with saturated sodiumbicarbonate solution (15 mL), brine (10 mL), dried (MgSO₄) andevaporated to give desired product 77H (121 m, 89%) as a white solid:R_(f) (10% MeOH/DCM) 0.39; HPLC (Phenomenex Luna 5 u C18 4.6×50 mm,linear gradient over 4 min) retention time 3.2 min (100%); MS (ES) m/z344.1 [M+H]⁺; HRMS calcd for C₂₀H₃₃N₃OSiCl 394.2081, Found 394.2080.

77I.4-[(8R,8aR)-8-(tert-Butyldimethylsilanyloxy)-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile(77Ia) and4-[(8S,8aS)-8-8-(tert-Butyldimethylsilanyloxy)-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile(77Ib)

To a solution of 77H (64 mg, 0.16 mmol) in DCM (2.8 mL) at 0° C. wasadded DIPEA (84 μL, 0.84 mmol, 3 equiv) followed by the addition ofphosgene (102 μL, 0.19 mmol, 1.2 equiv). The mixture was warmed to rtand stirred for 10 min. The reaction was concentrated and the residuewas purified by flash chromatography (0-50% EtOAc/hexane) to give theracemic mixtures of 77Ia and 77Ib (60 mg). A racemic mixture of 77Ia and77Ib (190 mg) was dissolved in IPA (5 mL) and purified by chiralpreparative HPLC (Chiralpak AD 4.6×250, 7% IPA/heptane) to giveenantiomer 77Ia (87 mg) and enantiomer 77Ib (85 mg). The relative andabsolute stereochemistry of 77Ia and 77Ib was assigned by separating theenantiomers of racemic intermediate 77C and transferring each enantiomerto the known (2S,3S) and (2R,3R)-3-hydroxypipecolic acids¹. Compounds77Ia and 77Ib were enantiomerically resynthesized from each enantiomersof intermediate 77C to confirm the stereochemistry. For 77Ia: Chiralanalytical HPLC (ChiralPak AD OD 4.6×250 mm, 20% IPA/hexane isocratic)retention time 11.0 min (100% ee); LRMS (ES⁺) [M+H]⁺ 420.1; HRMS calcdfor C₂₁H₃₁N₃O₂SiCl 420.1874, Found 420.1876. For 77Ib: Chiral analyticalHPLC (ChiralPak AD OD 4.6×250 mm, 20% IPA/hexane isocratic) retentiontime 15.6 min (100% ee); MS (ES) m/z 420.0 [M+1]⁺; HRMS calcd forC₂₁H₃lN₃O₂SiCl 420.1874, Found 420.1859.

77J.4-[(8R,8aR)-8-Hydroxy-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of 77Ia (11 mg, 0.026 mmol) in THF (1 mL) was added TBAF(1.0 M in THF, 1.5 mL). The reaction was stirred at rt for 2 h. Thereaction mixture was diluted with EtOAc, washed with water, dried(MgSO₄), filtered and concentrated to give the crude product.Purification by reverse phase HPLC (YMC-Pack ODS S-5 um 20×100 mm, 10min gradient, 20 mL/min, 20-100% solvent B, A=10% MeOH/water+0.1% TFA,B=90% MeOH/Water+0.1% TFA) provided the title compound 77J (7.1 mg):HPLC (Phenomenex Luna 5 u C18 4.6×50 mm, linear gradient over 4 min)retention time 2.30 min (100%); LC-MS [M+H]⁺ 306; Chiral analytical HPLC(ChiralPak AD OD 4.6×250 mm, 25% IPA/hexane isocratic) retention time13.9 min (100% ee); MS (ES) m/z 306.1 [M+1]⁺; HRMS calcd forC₁₅H₁₅N₃O₂Cl 304.0853, Found 304.0848.

77K.4-[(8S,8aS)-8-Hydroxy-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of 77Ib (11 mg, 0.026 mmol) in THF (1 mL) was added TBAF(1.0 M in THF, 1.5 mL). The reaction was stirred at rt for 2 h. Thereaction mixture was diluted with EtOAc, washed with water, dried(MgSO₄), filtered and concentrated to give the crude product.Purification by reverse phase HPLC (YMC-Pack ODS S-5 um 20×100 mm, 10min gradient, 20 mL/min, 20-100% solvent B, A=10% MeOH/water+0.1% TFA,B=90% MeOH/Water+0.1% TFA) provided the title compound (24 mg): HPLC(Phenomenex Luna 5 u C18 4.6×50 mm, linear gradient over 4 min)retention time 2.30 min (100%); LC-MS (M+H)⁺ 306; Chiral analytical HPLC(ChiralPak AD OD 4.6×250 mm, 25% IPA/hexane isocratic) retention time16.1 min (100% ee); MS (ES) m/z 306.1 [M+1]⁺; HRMS calcd forC₂₁H₃₁N₃O₂SiCl 304.0853, Found 304.0848. ² Lucia Battistini, FrancaZanardi et al, Tetrahedron: Asymmetry, 1997, 8, 2975-2987

EXAMPLE 784-[(8S,8aR)-8-Hydroxy-3-oxo-hexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile

78A.4-[(8aR)-3,8-Dioxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile

A mixture of oxalyl chloride (94 μL, 0.19 mmol) in DCM (50 μL) wascooled to −78° C. and a solution of DMSO (26 μL, 0.37 mmol) in DCM (0.7mL) was added. The resulting mixture was stirred at −78° C. for 15 min.A solution of4-[(8R,8aR)-8-hydroxy-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile(77J) (16 mg, 0.052 mmol) in DCM (0.5 mL) was followed by DCM (0.5 mL)and stirred at −78° C. for 30 min. Triethylamine (120 μL, 0.86 mmol) wasadded and gradually warmed to rt over 1 h. The solvent was removed andthe residue was purified by flash chromatography (silica gel, 30-50%EtOAc/hexane) to give the title compound 78A (12.9 mg) as a white solid:Chiral analytical HPLC (ChiralPak AD OD 4.6×250 mm, 35% IPA/hexaneisocratic) retention time 13.8 min (100% ee); MS (ES) m/z 304.1 [M+1]⁺;HRMS calcd for C₁₅H₁₃ClN₃O₂ 302.0696, Found 302.0681.

78B.4-[(8S,8aR)-8-Hydroxy-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile

A mixture of 78A (5 mg, 0.016 mmol) and sodium borohydride (5 mg) inMeOH:CH₃CN (1:1, 1.0 mL) was stirred at rt for 2 h. The reaction wasdiluted with EtOAc, washed with water (2×2 mL), dried (MgSO₄), filteredand concentrated to give the crude product. Purification by chiralpreparative HPLC (Chiralpak AD, 25% IPA/heptane isocratic) provided 78B(4 mg): HPLC (Phenomenex Luna 5 u C18 4.6×50 mm, linear gradient over 4min) retention time 2.40 min (100%); MS (ES) m/z 306 [M+1]⁺; Chiralanalytical HPLC (ChiralPak AD OD 4.6×250 mm, 25% IPA/hexane isocratic)retention time 12.1 min (97.6% ee); MS (ES) m/z 306.1 [M+1]⁺; HRMS calcdfor C₁₅H₁₅ClN₃O₂ 304.0853, Found 304.0846.

EXAMPLE 794-[(8R,8aS)-8-Hydroxy-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile

79A.4-[(8aS)-8-Hydroxy-3,8-dioxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile(20A)

Compound 79A was prepared using the same procedure as for intermediate78A from 77K (20 mg, 0.065 mmol), oxalyl chloride (2.0 M in DCM, 94 μL,0.19 mmol), DMSO (26 μL, 0.37 mmol), TEA (120 μL, 0.86 mmol) and DCM(0.55 mL) and purified by flash chromatography (4 g ISCO silica gelcolumn, 30-50% EtOAc/hexane) to give the title compound 79A (13 mg) as awhite solid: Chiral analytical HPLC (ChiralPak AD OD 4.6×250 mm, 35%IPA/hexane isocratic) retention time 11.3 min (64% ee).

79B.4-[(8R,8aS)-8-Hydroxy-3-oxohexahydroimidazo[1,5-a]pyridin-2-yl]-2-chloro-3-methylbenzonitrile

A mixture of 79A (13 mg, 0.033 mmol) and sodium borohydride (11 mg) inMeOH:CH3CN (1:1, 1.0 mL) was stirred at rt overnight. The reaction wasdiluted with EtOAc, washed with water (2×2 mL), dried (MgSO₄), filteredand concentrated to give the crude product (13 mg). Purification bychiral preparative HPLC (Chiralpak AD, 25% IPA/heptane isocratic)provided the title compound (4.5 mg): HPLC (Phenomenex Luna 5 u C184.6×50 mm, linear gradient over 4 min) retention time 2.40 min (100%);LC-MS [M+H]⁺ 306; Chiral analytical HPLC (ChiralPak AD OD 4.6×250 mm,25% IPA/hexane isocratic) retention time 10.5 min (98% ee); MS (ES): m/z306.1 [M+1]⁺; HRMS calcd for C₁₅H₁₅ClN₃O₂ 304.0853, Found 304.0863.

EXAMPLE 804-[(1S,7R,7aR)-7-Hydroxy-1-methyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

80A.(2S,3S)-N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-((1S)-1-hydroxyethyl)pyrrolidine(80A-a) and(2S,3S)-N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-((1R)-1-hydroxyethyl)pyrrolidine(80A-b)

To a solution of intermediate(2S,3S)-N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-formylpyrrolidine(4.7 g, 14.26 mmol) in THF (70 mL) at −78° C. was added dropwise MeMgBr(3.0 M in THF, 23.8 mmol). The reaction was stirred at −78° C. for 3 hand TLC (20% EtOAc/hexane) showed complete reaction. The reaction wasquenched by the addition of HOAc (5.0 mL) at −78° C., warmed to rt anddiluted with EtOAc (20 mL). The organic layer was separated and theaqueous layer was extracted with EtOAc (2×30 mL). The combined organiclayers were washed with saturated NaHCO₃ solution and brine, dried(MgSO₄), filtered and concentrated to give the crude product.Purification by flash chromatography (120 g ISCO silica gel column,0-30% EtOAc/hexane) provided 80A-a (760 mg) and 80A-b(1.54 g) ascolorless oil. 80A-a: For (20% EtOAc/hexane) 0.29. 80A-b: For (20%EtOAc/hexane) 0.25.

80B.(2S,3S)-N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-((1S)-1-methanesulfonyloxyethyl)pyrrolidine

To a solution of 80A-a (728 mg, 2.10 mmol) in DCM (10 mL) at 0° C. wasadded DIPEA (1.46 mL, 8.4 mmol) and a catalytic amount of DMAP. Themixture was stirred at 0° C. for 20 min and warmed to rt and stirred atrt for 4 h. The reaction was diluted with EtOAc and washed a 1:1 mixtureof 1 N HCl and brine (2×5 mL), saturated sodium bicarbonate solution (5ml), brine (5 mL), dried (MgSO₄), filtered and concentrated to give thecrude product. Purification by chromatography (10 g ISCO silica gelcartridge, 8% EtOAc/hexane) provided 80B as a yellow oil (410 mg): MS(ES) m/z 324 [M−Boc+H]⁺.

80C.(2R,3S)-N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-((1R)-1-azidoethyl)pyrrolidine

A mixture of 80B (650 mg, 1.53 mmol) and sodium azide (597 mg, 9,18mmol) was heated at 80° C. for 2 days. The reaction was diluted withEtOAc (200 mL), washed with brine:water (1:1, 4×15 mL), dried (MgSO₄),filtered and concentrated to give the crude product. Purification bychromatography (40 g ISCO silica gel cartridge, 0-20% EtOAc/hexane)provided 80C as a colorless oil (410 mg). MS (ES) m/z 271 [M−Boc+H]⁺.

80D.(2S,3S)-N-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-((1R)-1-aminoethyl)pyrrolidine

A mixture of 80C (410 mg, 1.10 mmol) and 10% Pd/C (100 mg) in MeOH:EtOAc(2:1, 15 mL) was stirred under hydrogen atmosphere for 2 h. The reactionwas filtered through a pad of celite. The filtrate was concentrated andpurified by flash chromatography (10 g ISCO silica gel cartridge, 0-20%MeOH/DCM) to the title compound 80D (284 mg): MS (ES) m/z 345 [M+H]⁺.

80E.(2S,3S)-1-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-[(1R)-1-(3-chloro-4-cyano-2-methylphenylamino)ethyl]pyrrolidine

To a solution of 80D (137 mg, 0.40 mmol) in nitrogen degassed toluene (3mL) at rt was added 2-chloro-4-iodo-3-methyl-benzonitrile (122 mg, 0.44mmol), Cs₂CO₃ (260 mg, 0.80 mmol) and Pd₂(dba)₃ (7.3 mg, 0.008 mmol) and(S)-N,N-dimethyl-1-[(R)-2-(diphenyphosphino)ferrocenyl]ethylamine (11mg, 0.024 mmol). The reaction was degassed with nitrogen for 30 min,sealed and heated at 140° C. for 2 h under microwave conditions. Themixture was cool to rt, diluted with hexane and filtered through a layerof celite. The filtrate was evaporated to give the crude product.Purification by flash chromatography (10 g ISCO silica gel cartridge,0-20% EtOAc in hexane gradient) gave the title compound 80E (36 mg): MS(ES) m/z 345 [M+H]⁺; HRMS calcd for C₂₅H₄₁N₃O₃ClSi 494.2606, found494.2606.

80F.(2R,3S)-3-(tert-Butyldimethylsilanyloxy)-2-[(1R)-1-(3-chloro-4-cyano-2-methylphenylamino)ethyl]pyrrolidine

Intermediate 80E (36 mg, 0.073 mmol) was dissolved in DCM (900 μL) andTFA (100 μL) was added. The mixture was stirred at rt for 10 h. Thereaction was diluted with EtOAc, washed with saturated sodiumbicarbonate solution (3×5 mL), brine (5 mL), dried (MgSO₄), filtered andconcentrated to give the title compound 80F (27 mg): MS (ES) m/z 394.5[M+H]⁺.

80G.4-[(1R,7S,7aR)-7-tert-Butyldimethylsilanoxy-1-methyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of crude 80F (˜0.068 mmol) in CH₂Cl₂ (2 mL) was addedi-Pr₂NEt (36 μL, 0.20 mmol) followed by 20% phosgene in toluene (50 μL,0.1 mmol)-dropwise. The reaction was stirred at rt for 4 days and nodesired product was formed. The solvent was removed and the residue wasdissolved in 1,2-dichloroethane (2 mL) and treated with i-Pr₂NEt (108μL) at 80° C. for 3 h. Reaction was concentrated and the residue waspurified by preparative TLC (30% EtOAc/hexane) to provide the titlecompound 80G (11 mg): MS (ES) m/z 420.5 [M+H]⁺; HRMS calcd forC₂₁H₃₁N3O₂ClSi 420.1874, found 420.1870.

80H.4-[(1S,7R,7aR)-7-Hydroxy-1-methyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of 80G (11 mg, 0.026 mmol) in THF (1 mL) was added TBAF(1.0 M in THF, 0.26 mL). The reaction was stirred at rt for 2 h. Thereaction mixture was concentrated and the residue was purified byreverse phase HPLC (Phenomenex Luna 20×100 mm S5 C18, 10 min. grad, 20mL/min, 20-100% B solvent, A=10% MeOH/water+0.1% TFA, B=90%MeOH/Water+0.1% TFA) to provide the title compound (5.1 mg): HPLC(Phenomenex Luna 5 u C18 4.6×50 mm, linear gradient over 4 min)retention time 2.40 min (100%); Chiral analytical HPLC (ChiralPak AD4.6×250 mm, 20% IPA/hexane isocratic) retention time 18.6 min (100% ee);MS (ES) m/z 306.1 [M+H]⁺; HRMS calcd for C₁₅H₁₇ClN₃O₂ 306.1009, Found306.1010.

Examples 81–84 MS (ES) Example Structure m/z [M + H]⁺ 81

320.1 82

320.1 83

320.1 84

320.181A.(±)-(2R,3R)-1-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-[(1R)-1-(3-chloro-4-cyano-2-methylphenylamino)ethyl]piperidine(81A-a) and (±)(2R,3R)-1-tert-Butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-[(1S)-1-(3-chloro-4-cyano-2-methylphenylamino)ethyl]piperidine(81A-b)

Compounds 81A-a and 81A-b were synthesized from1-tert-butyloxycarbonyl-3-(tert-butyldimethylsilanyloxy)-2-formyl-piperidine(18F) following a procedure similar to that used for 80F to give amixture of 81A-a and 81A-b as the crude product. Purification andseparation by flash chromatography (12 g ISCO silica gel column, 0-20%EtOAc/hexane gradient) provided 81A-a as colorless film (28 mg) and81A-b (14 mg) as a colorless film. For 81A-a: HPLC (Phenomenex Luna 5 uC18 4.6×50 mm, linear gradient over 4 min) retention time 4.85 min(100%); MS (ES) m/z 508.0 [M+H]⁺; HRMS calcd for C₂₆H₄₃ClN₃O₃Si508.2762, Found 508.2761. For 81A-b: HPLC (Phenomenex Luna 5 u C184.6×50 mm, linear gradient over 4 min) retention time 4.95 min (100%);MS (ES) m/z 508.0 [M+H]⁺; HRMS calcd for C₂₆H₄₃ClN₃O₃Si 508.2762, Found508.2748.

81B. Examples 81-82 and 83-84 were prepared from 81A-a and 81A-b,respectively, following the methods from Example 80. Individualenantiomers were isolated using preperative HPLC (ChiralPak 5×50 cm,isocratic 20% IPA/hexane).

EXAMPLE 854-[(1S,7S,7aR)-1-Ethyl-7-hydroxy-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl)-2-chloro-3-methylbenzonitrile

The title compound was prepared in a manner similar to that described inExample 61. Analytical HPLC with YMC Combiscreen ODS-A C18 4.6×50 mmcolumn, 4 min gradient, 10% MeOH/90% H₂O/0.1% H₃PO₄ to 90% MeOH/10%H₂O/0.1% H₃PO₄; 1 min hold, 4 mL/min UV detection at 220 nm, 98% purity(Rt 2.86 min); MS (ESI) m/z 320 [M+H]; HRMS calcd for [M−H] 318.1009,found 318.0994.

EXAMPLE 864-[(1R,7S,7aR)-1-Ethyl-7-hydroxy-3-oxohexahydropyrrolo[2-c]imidazol-2-yl)-2-chloro-3-methylbenzonitrile

The title compound was prepared in a manner similar to that described inExample 80. Analytical HPLC with YMC Combiscreen ODS-A C18 4.6×50 mmcolumn, 4 min gradient, 10% MeOH/90% H₂O/0.1% H₃PO₄ to 90% MeOH/10%H₂O/0.1% H₃PO₄; 1 min hold, 4 mL/min UV detection at 220 nm, 98% purity(Rt 2.67 min); MS (ESI) m/z 320 [M+H]; HRMS calcd for [M+H] 320.1166,found 320.1165.

EXAMPLE 87 4-[(1S, 7R,7aR)-1-Ethyl-7-hydroxy-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

87A.4-[(1S,7R,7aR)-7-Benzoyloxy-1-ethyl-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

To a solution of compound 85 (51 mg, 0.16 mmol) in 2 mL THF were addedtriphenylphosphine (84 mg, 0.32 mmol), benzoic acid (39 mg, 0.32 mmol)followed by diisopropyl azodicarboxylate (65 mg, 0.32 mmol) at rt. Themixture was stirred at rt for 2 h. Concentration of the reaction mixturevia rotary evaporator, followed by silica gel column chromatographypurification yielded the title compound (64 mg).87B.4-[(1S,7R,7aR)-1-Ethyl-7-hydroxy-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

Compound 87A (64 mg, 0.15 mmol) was dissolved in 1.5 mL of THF, treatedwith 1N KOH in anhydrous methanol (0.32 mL, 0.32 mmol) at rt for 2 h.Diluted the reaction mixture with 30 mL EtOAc, washed with saturatedaqueous NaHCO₃ solution (10 mL), brine (10 mL×2), dried over MgSO₄,filtered and concentrated. The resulting mixture was purified by reversephase HPLC (Phenominex Luna 30×250 mm S5 C18, 30 min. gradient time, 25mL/min, 30-100% B solvent, A=10% MeOH/water, B=90% MeOH/Water), providedthe title compound (43 mg). Analytical HPLC retention time 2.947 min(purity 98%). Condition: YMC Combiscreen ODS-A C18 4.6×50 mm column, 4min gradient, 4 mL/min flow, 0-100% B solvent, A=10% MeOH/water+0.1%H₃PO₄, B=90% MeOH/Water+0.1% H₃PO₄. MS (ES) m/z 320.24 [M+H]⁺, HRMScalcd for [M+H] 320.1166, found 320.1152.

EXAMPLE 884-[(1R,7R,7aR)-1-Ethyl-7-hydroxy-3-oxohexahydropyrrolo[1,2-c]imidazol-2-yl]-2-chloro-3-methylbenzonitrile

The title compound was prepared from compound 86 in a manner similar tothat described in Example 87. Analytical HPLC retention time 2.750 min(purity 96%). Condition: YMC Combiscreen ODS-A C18 4.6×50 mm column, 4min gradient, 4 mL/min flow, 0-100% B solvent, A=10% MeOH/water+0.1%H₃PO₄, B=90% MeOH/Water+0.1% H₃PO₄. MS (ES) m/z 320.22 [M+H]⁺, HRMScalcd for [M+H] 320.1166, found 320.1168.

EXAMPLE 892-Chloro-4-[(7R)-7-fluoro-3-oxotetrahydro-1H-pyrrolo[1,2-c]imidazol-2(3H)-yl]-3-methylbenzonitrile

A solution of(7S,7aS)-2-chloro-4-(7-hydroxy-3-oxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile(WO 03/096980) (30 mg, 0.103 mmol) in dry dichloromethane (1.0 mL) wascooled to −65° C., treated with DAST (26 μL, 0.21 mmol) and stirred for17 h, allowing the temperature to warm to rt. The reaction mixture wasthen cooled to 0° C., quenched with 10% sodium carbonate (0.5 mL) andextracted with CH₂Cl₂ (2×10 mL). The organic phase was washed with H₂O(0.5 mL), dried (Na₂SO₄), filtered and concentrated under reducedpressure. The syrup was combined with a previous batch (5.4 mg) andchromatographed 3× (silica gel; EtOAc/hexane gradient then CH₃OH/CH₂Cl₂gradient) to yield the title compound (14.5 mg, 47.9%). HPLC: 99.1% at2.04 min (retention time) (Conditions: YMC S-5 C-18 (4.6×50 mm), elutingwith 0-100% B, 4 min gradient. (A=90% H₂O—10% CH₃C—0.1% TFA and B=10%H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4 mL/min. UV detection at 220 nm).Chiral HPLC: retention time=20.8 min (100%); Conditions: OD (4.6×250mm); Eluted with 20% isopropanol in heptane for 60 min at 1 mL/min. MS(ES⁺): m/z 294 [M+H]⁺.

EXAMPLE 904-[(7R)-7-Amino-3-oxotetrahydro-1H-pyrrolo[1,2-c]imidazol-2(3H)-yl]-2-chloro-3-methylbenzonitrile

90A.(7S)-2-(3-Chloro-4-cyano-2-methylphenyl)-3-oxohexahydro-1H-pyrrolo[1,2-c]imidazol-7-ylmethanesulfonate

A cooled (0° C.) solution of(7S,7aS)-2-chloro-4-(7-hydroxy-3-oxotetrahydropyrrolo[1,2-c]imidazol-2-yl)-3-methylbenzonitrile(WO 03/096980) (342 mg, 1.17 mmol) and triethylamine (0.70 mL, 5.02mmol) in dry CH₂Cl₂ (7.0 mL) was treated with methanesulfonyl chloride(0.21 mL, 2.71 mmol) and stirred, allowing the temperature to come up tort over 17 h. The mixture was diluted with CH₂Cl₂ (100 mL), washed withsaturated NaHCO₃ (4.0 mL) and brine (4.0 mL), dried (Na₂SO₄), filteredand concentrated under reduced pressure. The syrup was chromatographed(silica gel; CH₂Cl₂/CH₃OH gradient) to yield the title compound (400.2mg, 92.5%) as a white solid foam from CH₂Cl₂/Hexane. HPLC: 97.9% at 2.10min (retention time) (Conditions: YMC S-5 C-18 (4.6×50 mm), eluting with0-100% B, 4 min gradient. (A=90% H₂O—10% CH₃CN—0.1% TFA and B=10%H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4 mL/min. UV detection at 220 nm.MS (ES⁺): m/z 370 [M+H]⁺.

90B.2-Chloro-4-[(7R,7aR)-7-azido-3-oxotetrahydro-1H-pyrrolo[1,2-c]imidazol-2(3H)-yl]-3-methylbenzonitrile

A solution of compound 90A (400 mg, 1.18 mmol) and sodium azide (309 mg,4.75 mmol) in anhydrous DMF (3.2 mL) was placed in a pre-heated bath(75° C.) and stirred at 75° C. for 3 h. The solution was cooled,quenched with H₂O (6.0 mL) and extracted with EtOAc (2×80 mL). Theorganic phase was washed with H₂O (6.0 mL), brine (6.0 mL), dried(Na₂SO₄), filtered and concentrated under reduced pressure. The syrupwas chromatographed (silica gel; CH₂Cl₂/CH₃OH gradient) to yield thetitle compound (339.3 mg, 90.8%) as a white solid foam fromCH₂Cl₂/Hexane. HPLC: 97.7% at 2.22 min (retention time) (Conditions: YMCS-5 C-18 (4.6×50 mm), eluting with 0-100% B, 4 min gradient. (A=90%H₂O—10% CH₃CN—0.1% TFA and B=10% H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4mL/min. UV detection at 220 nm. MS (ES⁺): m/z 317 [M+H]⁺.

90C.4-[(7R)-7-Amino-3-oxotetrahydro-1H-pyrrolo[1,2-c]imidazol-2(3H)-yl]-2-chloro-3-methylbenzonitrile

A solution of compound 90B (20.5 mg, 0.06 mmol) in dry MeOH (1.0 mL) wastreated with PtO₂ (5.5 mg) and hydrogenated (balloon) at rt for 1.0 h.The reaction mixture was diluted with MeOH (1.0 mL) and filtered througha syringe filter, washing the syringe filter well with MeOH (3×1.0 mL).The clear filtrate was concentrated under reduced pressure and theresidue was chromatographed (silica gel; CH₂Cl₂/CH₃OH gradient) to yieldthe title compound (11.9 mg, 68.2%) as a white solid. HPLC: 100% at 1.38min (retention time) (Conditions: YMC S-5 C-18 (4.6×50 mm), eluting with0-100% B, 4 min gradient. (A=90% H₂O—10% CH₃CN—0.1% TFA and B=10%H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4 mL/min. UV detection at 220 nm.Chiral HPLC: retention time=18.98 min (99.6%); Conditions: OD (4.6×250mm); Eluted with 20% isopropanol in heptane for 30 min at 1 mL/min. MS(ES⁺): m/z 291 [M+H]⁺.

EXAMPLE 912-Chloro-3-methyl-4-[(7R)-7-methylamino-3-oxotetrahydro-1H-pyrrolo[1,2-c]imidazo-2(3H)-yl]benzonitrile

A solution of compound 90B (32 mg, 0.10 mmol) and 1.0 M Et₃P in THF(0.23 mL, 0.23 mmol) in anhydrous CH₂Cl₂ (0.85 mL) was stirred at rt for1.5 h, diluted with dry CH₂Cl₂ (0.85 mL) then treated with methyl iodide(63 μL, 1.0 mmol). The reaction mixture was stirred at rt for 3.0 h,diluted with CH₂Cl₂ (3.0 mL), treated with H₂O (0.5 mL) and stirred atrt for another 30 min. The reaction mixture was extracted with CH₂Cl₂(2×10 mL), dried over Na₂SO₄, filtered and concentrated under reducedpressure. The resulting syrup was chromatographed (silica gel;CH₂Cl₂/CH₃OH gradient) followed by preparative HPLC and trituration ofthe product obtained with Et₂O (2×0.5 mL) to yield the title compound(8.2 mg, 19.6%) as a light beige solid. HPLC: 98.0% at 1.35 min(retention time) (Conditions: YMC S-5 C-18 (4.6×50 mm), eluting with0-100% B, 4 min gradient. (A=90% H₂O—10% CH₃CN—0.1% TFA and B=10%H₂O—90% CH₃CN—0.1% TFA); Flow rate at 4 mL/min. UV detection at 220 nm.MS (ES⁺): m/z 305 [M+H]⁺.

EXAMPLE 92N-(7R)-2-(3-Chloro-4-cyano-2-methylphenyl)-3-oxohexahydro-1-pyrrolo[1,2-c]imidazol-7-yl]-2,2,2-trifluoroacetamide

A solution of compound 90C (20 mg, 0.069 mmol) in anhydrous CH₂Cl₂ (0.50mL) was cooled to 0° C., treated with trifluoroacetic anhydride (13 μL,0.09 mmol), stirred at 0° C. for 1.0 h then at rt for 17 h. The reactionmixture was diluted with EtOAc (15 mL) and washed with 5% NaHCO₃ (0.5mL). The organic phase was dried over anhydrous Na₂SO₄, filtered,concentrated under reduced pressure and combined with another batch from91C (0.076 mmol). The syrup was chromatographed (silica gel;CH₂Cl₂/CH₃OH gradient) to yield the title compound (38 mg, 64.5%) as awhite solid foam from CH₂Cl₂/Hexane. HPLC: 97.0% at 2.1 min (retentiontime) (Conditions: YMC S-5 C-18 (4.6×50 mm), eluting with 0-100% B, 4min gradient. (A=90% H₂O−10% CH₃CN—0.1% TFA and B=10% H₂O−90% CH₃CN−0.1%TFA); Flow rate at 4 mL/min. UV detection at 220 nm. Chiral HPLC:retention time=10.78 min (98.2%); Conditions: OD (4.6×250 mm); Elutedwith 20% isopropanol in heptane for 30 min at 1 mL/min. MS (ES+): m/z387 [M+H]⁺.

1. A compound according to formula I

or a pharmaceutically acceptable salt thereof wherein R₁ is selectedfrom hydrogen (H), alkenyl or substituted alkenyl, CO₂R₄, CONR₄R₄′ andCH₂OR₄; R₂ and R₂′ are each independently selected from hydrogen (H),alkyl, substituted alkyl, SR₃, halo, NHR₄, NHCOR₄, NHCO₂R₄, NHCONR₄R₄′and NHSO₂R₄; and at least one of R₂ and R₂′ is H or alkyl; R₃ in eachfunctional group is independently selected from hydrogen (H), alkyl orsubstituted alkyl, CHF₂, CF₃ and COR₄; R₄ and R₄′ in each functionalgroup are each independently selected from hydrogen(H), alkyl orsubstituted alkyl, alkenyl or substituted alkenyl, alkynyl orsubstituted alkynyl, cycloalkyl or substituted cycloalkyl, arylalkyl orsubstituted arylalkyl, and aryl or substituted aryl; R₅ and R₅′ are eachindependently selected from hydrogen(H), alkyl or substituted alkyl,alkenyl or substituted alkenyl, alkynyl or substituted alkynyl,cycloalkyl or substituted cycloalkyl and arylalkyl or substitutedarylalkyl, wherein at least one of R₅ and R₅′ is hydrogen, or R₅ and R₅′taken together can form a double bond with oxygen (O), sulfur (S), NR₇or CR₇R₇′; R₆ and R₆′ are each independently at each occurrence selectedfrom hydrogen(H), alkyl or substituted alkyl, alkenyl or substitutedalkenyl, alkynyl or substituted alkynyl, cycloalkyl or substitutedcycloalkyl, and arylalkyl or substituted arylalkyl, wherein at least oneof R₆ and R₆′ is hydrogen, or R₆ and R₆′ at each occurrence takentogether form a double bond with oxygen (O), sulfur (S), or CR₇R₇′; R₇and R₇′ in each functional group are each independently selected fromhydrogen(H), OR₄, alkyl or substituted alkyl, alkenyl or substitutedalkenyl, alkynyl or substituted alkynyl, cycloalkyl or substitutedcycloalkyl, arylalkyl or substituted arylalkyl, and aryl or substitutedaryl; G is an aryl group, wherein said group is mono- or polycyclic, andwhich is optionally substituted with one or more substitutents selectedfrom hydrogen, halo, CN, CF₃, OR₄, CO₂R₄, NR₄R₄′, CONR₄R₄′, CH₂OR₄,alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl orsubstituted alkynyl, cycloalkyl or substituted cycloalkyl, arylalkyl orsubstituted arylalkyl, and aryl or substituted aryl; W is selected from(CR₆R₆′), C(R₆)OR₃, or C(R₆)(NR₄R₄′); and n is 1; with the followingprovisos: (a) when R₅ and R₅′ and/or R₆ and R₆′ form a double bond withCR₇R₇′, then when either R₇ or R₇′ is OR₄, R₄ is not hydrogen; (b) when(i) R₅ and R₅′ are each H or taken together to be =O, =S or =CH₂, (ii)R₆ and R₆′ on the imidazolidine portion of the bicyclic structure shownare selected from hydrogen(H), alkyl or substituted alkyl, alkenyl, orsubstituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl orsubstituted cycloalkyl, and arylalkyl or substituted aryalkyl, whereinat least one of R₆ and R₆′ on the imidazolidine portion of the bicyclicstructure shown is hydrogen, or R₆ and R₆′ on the imidazolidine portionof the bicyclic structure shown are taken together to form a double bondwith oxygen (O) or sulfur (S), (iii) W is CR₆R₆′ where R₆ and R₆′ areeach independently selected from H, alkyl or substituted alkyl, alkenylor substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl orsubstituted cycloalkyl and arylalkyl or substituted arylalkyl, whereinat least one of R₆ and R₆′ in W is H, and (iv) G has the structure:

wherein, R₁₃ is selected from the group consistng of H, CN, NO₂, halo,OR₁₄, CO₂R₁₅, CONHR₁₅, COR₁₅, S(O)_(p)R₁₅, SO₂NR₁₅R₁₅′, NHCOR₁₅ andNHSO₂R₁₅, wherein p is an integer from 0 and 2, R₁₄ in each functionalgroup is independently selected from H, alkyl or substituted alkyl,CHF₂, CF₃ and COR₁₅, R₁₅ and R₁₅′ in each functional group are eachindependently selected from H, alkyl or substituted alkyl, alkenyl orsubstituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl orsubstituted cycloalkyl, arylalkyl or substituted arylalkyl, aryl orsubstituted aryl, heteroaryl or substituted heteroaryl and -CN, and Aand B are each independently selected from H, halo, CN, NO₂, alkyl orsubstituted alkyl and OR₁₄, then R₂ R₂′ are each independently selectedfrom SR₃ and NHR₄.
 2. The compound or pharmaceutically acceptable saltthereof according to claim 1 wherein G is selected from:

wherein R₈, R₉, R₁₀ and R₁₁ are each independently selected fromhydrogen (H), NO₂, CN, CF₃, OR₄, CO₂R₄, NR₄R₄′, CONR₄R₄′, CH₂OR₄, alkylor substituted alkyl, alkenyl or substituted alkenyl, alkynyl orsubstituted alkynyl, cycloalkyl or substituted cycloalkyl, arylalkyl orsubstituted arylalkyl, and aryl or substituted aryl; A to F is eachindependently selected from N or CR₉; J, K, L, P and Q are eachindependently selected from CR₁₂R₁₂′; R₁₂ and R₁₂′ in each functionalgroup are each independently selected from a bond or R₁; and m is aninteger of 0 or
 1. 3. The compound or pharmaceutically acceptable saltthereof according to claim 2 wherein R₈ is -CN.
 4. The compoundaccording to claim 1 which is:

or a pharmaceutically acceptable salt thereof.
 5. The compound accordingto claim 1 selected from:

or a pharmaceutically acceptable salt thereof.
 6. A pharmaceuticalcomposition, comprising: (a) a compound or pharmaceutically acceptablesalt thereof according to claim 1; and (b) at least one pharmaceuticallyacceptable diluent or carrier.
 7. The compound according to claim 1which is

or a pharmaceutically acceptable salt thereof.
 8. The compound accordingto claim 1 which is

or a pharmaceutically acceptable salt thereof.
 9. A pharmaceuticalcomposition according to claim 6, wherein the compound according toclaim 1 is

or a pharmaceutically acceptable salt thereof.
 10. A pharmaceuticalcomposition according to claim 6, wherein the compound according toclaim 1 is

or a pharmaceutically acceptable salt thereof.
 11. The compoundaccording to claim 1 which is

or a pharmaceutically acceptable salt thereof.
 12. The compoundaccording to claim 1 which is

pharmaceutically acceptable salt thereof.
 13. The compound according toclaim 1 which is

pharmaceutically acceptable salt thereof.
 14. The compound according toclaim 1 which is

pharmaceutically acceptable salt thereof.
 15. The compound according toclaim 1 which is

pharmaceutically acceptable salt thereof.
 16. The compound according toclaim 1 which is

pharmaceutically acceptable salt thereof.
 17. The compound according toclaim 1 which is

pharmaceutically acceptable salt thereof.
 18. The compound according toclaim 1 which is

pharmaceutically acceptable salt thereof.
 19. The compound according toclaim 1 which is

pharmaceutically acceptable salt thereof.
 20. The compound according toclaim 1 which is

pharmaceutically acceptable salt thereof.
 21. The compound according toclaim 1 which is

a pharmaceutically acceptable salt thereof.
 22. The compound accordingto claim 1 which is

a pharmaceutically acceptable salt thereof.